Registration Dossier

Administrative data

Description of key information

Under the conditions of this study, this substance was not found to be an irritant or corrosive.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records
Reference
Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
4 July 2018 (Experimental Start) to 6 July 2018 (Experimental Completion)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)
Deviations:
no
Qualifier:
according to
Guideline:
other: Commission Regulation (EC) No 440/2008, Annex Part B, B.40.Bis: “In Vitro Skin Corrosion: Human Skin Model Test”, Official Journal of the European Union No. L142 (31 May 2008)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Materia Inc.
- Lot/batch No.of test material: RR229-0714
- Expiration date of the lot/batch: 6 June 2020
- Purity test date: 28 March 2018

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25 ºC, below 70 RH%), under inert (Nitrogen) gas
- Stability under test conditions: Assumed stable for the duration of the test
- Solubility and stability of the test substance in the solvent/vehicle: The test item was applied as supplied, no formulation required.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: None, administered to the test system as supplied

OTHER SPECIFICS:
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is added: Not specified, test item applied to the test system as supplied.
Test system:
human skin model
Remarks:
EPISKIN TM(SM)
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Adult human derived epidermal keratinocytes (EPISKIN TM(SM) (Manufacturer: SkinEthic, France, Batch No.: 18-EKIN-027, Expiry Date: 09 July 2018
Source strain:
other: Strain No's: 10-KERA-003 & 10-KERA-004
Justification for test system used:
The EPISKIN TM (SM) model has been validated for corrosivity and irritation testing in an international validation study and its use is recommended by the relevant OECD guidelines for corrosivity and irritation testing (OECD No. 431 and OECD No. 439). It was therefore considered as suitable for use in this study.
Vehicle:
unchanged (no vehicle)
Remarks:
Test item was applied as supplied, no formulation required.
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EPISKIN TM (SM) Model
- Tissue batch number: EPISKIN TM(SM) (Manufacturer: SkinEthic, France, Batch No.: 18-EKIN-027, Expiry Date: 09 July 2018)
- Production date: 3 July 2018
- Expiry date: 9 July 2019
- Shipping date: Not specified
- Delivery date: Not specified
- Date of initiation of testing: 4 July 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: Disks of EPISKIN TM(SM) were treated with the test item and incubated for 4 hours (corrosivity testing) and for 15 minutes (irritation testing) at room temperature.
- Temperature of post-treatment incubation (if applicable): After 15 minutes incubation time (irritation test) or 4 hours incubation time (corrosivity test), the EPISKIN TM (SM) units were removed and rinsed thoroughly with PBS to remove any remaining material from the epidermal surface. The rest of the PBS was removed from the epidermal surface with a pipette (without touching the epidermis).

For the irritation test, after rinsing, the units were placed into the plate wells with fresh pre-warmedMaintenance Medium (2 mL/well) below them and then incubated for 42 hours (±1 hour) at 37 °C in an incubator with 5% CO2, in a > 95% humidified atmosphere.

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: After 15 minutes incubation time (irritation test) or 4 hours incubation time (corrosivity test), the EPISKINTM (SM) units were removed and rinsed thoroughly with PBS to remove any remaining material from the epidermal surface. The rest of the PBS was removed from the epidermal surface with a pipette (without touching the epidermis).
- Observable damage in the tissue due to washing: None
- Modifications to validated SOP: None

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: In both the irritation and corrosion tests, MTT solution (2 mL of 0.3 mg/mL MTT working solution) was added to each well below the skin units (except for the two living colour control units). The lid was replaced and the plate incubated at 37 °C in an incubator with 5% CO2 for 3 hours (±5 minutes), protected from light, in a > 95% humidified atmosphere.
- Incubation time: 37 °C in 5% CO2 for 3 hours (±5 minutes), protected from light, in a > 95% humidified atmosphere.
- Spectrophotometer: Not specified
- Wavelength: The OD (optical density or absorbance) of samples was measured using a plate reader at 570 nm
- Filter: Not specified
- Filter bandwidth: Not specified
- Linear OD range of spectrophotometer: Not specified

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: In the irritation test, the mean cell viability was 105.6% compared to the negative control. This is above the threshold of 50%, therefore the test item was considered as non-irritant to skin.
- Viability: In the corrosivity test, the mean cell viability was 109.6% compared to the negative control. This is above the threshold of 35%, therefore the test item was considered as being non-corrosive.
- Barrier function: Appropriate cell morphology. The EPISKIN TM(SM) kits are manufactured according to ISO 9001. All biological components of the epidermis and the
kit culture medium have been tested for the presence of viruses, bacteria and mycoplasma. The quality of the final product is assessed by undertaking a MTT cell
viability test and a cytotoxicity test with sodium dodecyl sulphate (SDS). These quality control experiments were conducted at SkinEthic laboratories (supplier of the
EPISKIN TM(SM) Test Kits used in this study.
- Morphology: A highly differentiated and stratified epidermis model is obtained after 13-day culture period comprising the main basal, supra basal, spinous and granular layers and a functional stratum corneum (Tinois et al., 1994)
- Contamination: None
- Reproducibility: All parameters were within acceptable limits and therefore the study was considered as valid.

NUMBER OF REPLICATE TISSUES:
In the corrosivity test, two replicates per time point were used for the test item accompanied by two negative controls and two positive controls.
In the irration test, three replicates per time point were used for the test item accompanied by three negative controls and three positive controls.
For both tests, two additional test item-treated living tissues were used for the non-specific OD evaluation.

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues : For both tests, two additional test item-treated living tissues were used for the non-specific OD evaluation. No killed tissues were used.
- N. of replicates: two
After three hours of incubation, a yellow colour of the mixture was detected in the test tube. The test item did not therefore react with MTT and thus the use of additional controls was not necessary.

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

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be non-corrosive to skin if following exposure with Distilled Tricyclopentadiene, the mean cell viability was above the threshold of 35%. As the mean cell viability was 109.6% following exposure to TCPD when compared to the negative control, the test item was therefore considered as non-corrosive.

- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439:
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
Negative Control

Corrosivity testing:
Physiological saline (0.9% (w/v) NaCl solution):

Manufacturer: B. Braun Pharmaceuticals SA
Batch No.: 80353Y05-1
Expiry Date: December 2020
Grade: Sterile


Irritation testing:

Phosphate Buffered Saline:
Abbreviation: PBS
Supplier: Life Technologies Corporation
Batch number: 1930056
Expiry date: November 2019


Positive Control

Corrosivity testing:
Glacial acetic acid:
Supplier: VWR International Ltd.
Batch No.: 17H074111
Expiry date: 05 July 2020


Irritation testing:

5% (w/v) Sodium Dodecyl Sulphate solution:
The positive control solution (abbreviated as SDS in the raw data and report) was prepared freshly in the testing laboratory. The following chemicals were used for the preparation of the positive control solution:

Sodium Dodecyl Sulphate:

Supplier: REANAL
Batch number: PP/2016/11637
Expiry date: 30 June 2021

Distilled water:
Supplier: B. Braun Pharmaceuticals SA
Batch number: 73751Y25-2
Expiry date: August 2020


Duration of treatment / exposure:
In case of the corrosivity testing, 30 mg of test item was applied evenly to each of two test units and each additional control skin units and then 100 µL physiological saline was added to the test item to ensure good contact with the epidermis in each case. In case of the irritation testing, first an appropriate amount (10 µL) distilled water was applied to the epidermal surface in order to improve further contact between test item and epidermis and then 30 mg of test item were applied evenly to each of three test units and each additional control skin units. The amounts were sufficient to cover the epidermal surface.
Duration of post-treatment incubation (if applicable):
After 15 minutes incubation time (in the irritation test) or 4 hours incubation time (in the corrosivity test), the EPISKINTM (SM) units were removed and rinsed thoroughly with PBS to remove any remaining material from the epidermal surface as much as possible. The rest of the PBS was removed from the epidermal surface with a pipette (without touching the epidermis).
Number of replicates:
Two replicates per time point were used for test item (corrosivity test). Two negative controls and two positive controls were also run in the corrosivity test.
Three replicates per time point were used for test item (irritation test). Three negative controls and three positive controls were also run in irritation test.
In both tests, two additional test item-treated living tissues were used for the non-specific OD evaluation.
Irritation / corrosion parameter:
% tissue viability
Remarks:
Corrosivity test
Run / experiment:
1 (Negative control) Physiological saline
Value:
100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
No indication of corrosivity
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 (Test item - TCPD)
Value:
109.6
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
No indication of corrosivity
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1 (Positive control) Glacial acetic acid
Value:
0.6
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other:
Remarks:
No indication of corrosivity
Other effects / acceptance of results:
- OTHER EFFECTS:
- Visible damage on test system: None
- Direct-MTT reduction: After three hours of incubation the test item did not react with MTT.
- Colour interference with MTT: No colour change was observed after three hours incubation of the test item in MTT working solution, the test material did not interact with MTT. Therefore, additional controls and data calculations were not necessary. The false estimation of viability can be excluded.

DEMONSTRATION OF TECHNICAL PROFICIENCY: The test facility chosen to conduct these tests (OECD 431 and OECD 439) is technically proficient in these in vitro tests, is GLP compliant and has appropriate historical data on which to compare the test item againsts to define a study outcome of non-corrosive and non-irritant to skin.

ACCEPTANCE OF RESULTS:
General validity criteria: The mean OD value of the two or three negative control tissues should be ≥ 0.6 and ≤ 1.5. The mean OD value of the blank samples (acidified isopropanol) should be < 0.1.

- Acceptance criteria met for negative control:
In the corrosivity test, the mean OD value of the two negative control tissues was in the recommended range (0.830).
In the irritation test, the mean OD value of the three negative control tissues was in the recommended range (0.876).

- Acceptance criteria met for positive control:
In the corrosivity test, the two positive control treated tissues showed 0.6% viability demonstrating the proper performance of the assay.
In the irritation test, the positive control treated tissues showed 1.6% viability demonstrating the proper performance of the assay.

- Acceptance criteria met for variability between replicate measurements:
In the corrosivity test, the difference of viability between the two test item-treated tissue samples in the MTT assay was 4.6%. The difference of viability between the two negative control tissue samples in the MTT assay was 3.8%.

In the irritation test, the standard deviation of the viability results for negative control samples was 3.8%. The standard deviation of viability values of the three test item-treated tissue samples in the MTT assay was 7.5%.

All parameters were within acceptable limits and therefore the studies considered valid.
Interpretation of results:
GHS criteria not met
Conclusions:
In this in vitro EPISKIN TM (SM) model test with Distilled Tricyclopentadiene (TCPD), the results indicate that the test item is non-corrosive to the skin, UN GHS Classification: No Category.
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:
16 August 2018 (Experimental Start) to 22 January 2019 (Experimental Completion)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)
Version / remarks:
none
Deviations:
no
GLP compliance:
yes (incl. certificate)
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source of test material: Materia Inc.
- Lot/batch No.of test material: RP229-0714
- Expiration date of the lot/batch: 06 June 2020
- Purity test date: 28 March 2018 (CoA)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Controlled room temperature (15-25 ºC, below 70% Relative Humidity), under inert (N2) gas.
- Stability under test conditions: Assumed stable for the duration of the test
- Solubility and stability of the test substance in the solvent/vehicle: Non-soluble in physiological saline. The test item was applied in its original form, as supplied.
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: Not applicable.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: No, test item applied in its original form, as supplied.

OTHER SPECIFICS:
- measurement of pH, osmolality, and precipitate in the culture medium to which the test chemical is added: Not specified
Species:
chicken
Strain:
other: Ross 308
Remarks:
Source: TARAVIS KFT. 9600 Sárvár, Rábasömjéni út. 129., Hungary
Vehicle:
unchanged (no vehicle)
Remarks:
Test item applied in its original form.
Controls:
yes, concurrent positive control
yes, concurrent negative control
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): In each experiment, as the test item was waxy, an alternative treatment method was used: a thin even layer of test item (ca. 1 g) was spread on a disk of Parafilm and then placed onto the cornea surface to ensure suitable exposure. The amount of test item was enough to adequately expose the entire surface of the cornea.

- In each experiment one negative control eye was treated with 30 µL of physiological saline. An additional negative control eye (“Parafilm control”) was treated using the alternative treatment method to show it had no impact on the results.
- In each experiment positive control eyes were treated with 30 mg powdered Imidazole.
Duration of treatment / exposure:
10 seconds
Duration of post- treatment incubation (in vitro):
The time of application was noted, then after an exposure period of 10 seconds from the end of application the cornea surface was rinsed thoroughly with 20 mL physiological saline solution at ambient temperature, taking care not to damage the cornea but attempting to remove all residual test material.

Additional gentle rinsing with at least 20 mL saline was performed at each time point (30, 75, 120, 180 and 240) when the test item or positive control material remaining on the cornea was observed.
Number of animals or in vitro replicates:
In each experiment, three test item treated eyes, three positive control treated eyes, one negative control and one parafilm control treated eye were examined.
Details on study design:
SELECTION AND PREPARATION OF ISOLATED EYES
Eyes selection: On receipt the chicken head was placed on soft paper. The eyelids were carefully cut away with scissors, avoiding damaging the cornea. One small drop of 2% (w/v) fluorescein solution was applied onto the cornea surface for a few seconds and subsequently rinsed off with 20 mL physiological saline. The fluorescein-treated cornea was examined with a hand-held slit lamp or slit lamp microscope, to ensure that the cornea was not damaged. If the cornea was in good condition, the eyeball was carefully removed from the orbit.

Preparation of eyes: The eye ball was carefully removed from the orbit by holding the nictitating membrane with surgical forceps, while cutting the eye muscles with bent scissors. Care was taken to remove the eyeball from the orbit without cutting the optical nerve too short. The procedure avoided pressure on the eye while removing the eyeball from the orbit to prevent distortion of the cornea and subsequent corneal opacity. Once removed, the eye was placed onto damp paper and the nictitating membrane cut away with other connective tissue. The prepared eyes were kept on wet papers in a closed box so appropriate humidity was maintained.

The prepared eye was placed in a steel clamp, avoiding too much pressure on the eye, with the cornea positioned vertically (i.e. in the same position as in the chicken head). Due to the relatively firm sclera of the chicken eyeball, only slight pressure was needed to fix the eye properly. The clamp with the eyeball was transferred to a chamber of the superfusion apparatus. The clamp holding the eye was positioned in such a way that the entire cornea was supplied with physiological saline solution dripping from a stainless steel tube (at a rate of approximately 3-4 drops/minute or 0.1 to 0.15 mL/minutes). The chamber door was closed with the exception of any subsequent manipulation or examinations, to maintain temperature and humidity.

Eyes examination and acclimatization time: The appropriate number of eyes were selected and placed in the superfusion apparatus. There they were examined again with the slit lamp microscope to ensure they were in good condition. The focus was adjusted to clearly see the physiological saline flowing on the cornea surface. Eyes with a high baseline fluorescein staining (i.e., > 0.5) or corneal opacity score (i.e., > 0.5) were rejected. The cornea thickness was measured, any eye with cornea thickness deviating more than 10 % from the mean value for all eyes, or eyes that showed any other signs of damage, were rejected and replaced. If the selected eyes were appropriate for the test, then acclimatization was started and conducted for approximately 45 to 60 minutes. The chambers of the superfusion apparatus were at a controlled temperature (32 ±1.5°C) during the acclimatization and treatment periods.

EQUILIBRATION AND BASELINE RECORDINGS
At the end of the acclimatization period, a zero reference measurement was recorded for cornea thickness and opacity to serve as a baseline (t=0) for each individual eye. The cornea thickness should not change by more than 5% within the -45 min and the zero time. No changes in thickness (0.0%) were observed in the eyes in experiments. Following the equilibration period, the fluorescein retention was measured. Baseline values were required to evaluate any potential test item related effect after treatment. All eyes were considered suitable for assay use.

NUMBER OF REPLICATES : In each experiment, three test item treated eyes, three positive control treated eyes, one negative control and one "parafilm control" treated eye were examined.

NEGATIVE CONTROL USED : 30 µL of physiological saline (0.9% (w/v) NaCl solution)

An additional control eye (i.e. parafilm control, using one eye) was also run in conjunction with the test item to show that the alternative treatment method (as the test item was waxy, due to the physical nature of the test item an alternative treatment method of treatment was used) had no impact on the results.

POSITIVE CONTROL USED : 30 mg powdered Imidazole.

APPLICATION DOSE AND EXPOSURE TIME : Ca.1g of test item was applied onto the centre of the cornea in such a way that the entire surface of the cornea was uniformly covered (due to the physical nature of the test item an alternative treatment method was used). After 10 seconds, the surface was rinsed with 20 mL physiological saline solution at ambient temperature.

OBSERVATION PERIOD : The control eyes and test item treated eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse.

REMOVAL OF TEST SUBSTANCE
- Volume and washing procedure after exposure period: Post exposure and rinseing, an additional gentle rinsing with at least 20 mL saline was performed at each time point (30, 75, 120, 180 and 240) until all residual test item or positive control material on the cornea had been removed.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Corneal thickness and corneal opacity were measured at all time points. A Haag-Streit Bern 900 slit-lamp microscope was used for measurements.



Corneal swelling was calculated according to the following formulae:
CS (Corneal swelling) at time t = CT (Corneal thickness) at time t –CT at t=0 / CT at t=0 (x100)

Mean CS at time t = FECS(at time t) + SECS(at time t) + TECS(at time t) / 3

FECS(at time t) = first eye cornea swelling at a given time-point
SECS(at time t) = second eye cornea swelling at a given time-point
TECS(at time t) = third eye cornea swelling at a given time-point

- Damage to epithelium based on fluorescein retention: The cornea thickness was measured, any eye with cornea thickness deviating more than 10 % from the mean value for all eyes, or eyes that showed any other signs of damage, were rejected and replaced. Fluorescein retention was measured on two occasions, at baseline (t=0) and approximately 30 minutes after the post-treatment rinse. A Haag-Streit Bern 900 slit-lamp microscope was used for the measurements.


- Swelling: measured with optical pachymeter on a slit-lamp microscope; slit-width setting:
Small negative numbers for swelling (0 to -5%) following application are evaluated as class I.
Large negative numbers (>12% below control) are probably due to erosion and indicate a severe effect (scored as class IV).
Cases of values of -5% to -12% are evaluated on a case by case basis but in the absence of other findings do not indicate a severe effect (class II).


- Macroscopic morphological damage to the surface: None
- Mean maximum opacity score : See below tables for ICE classification criteria for corneal thickness and opacity:
- Mean fluorescein retention score at 30 minutes post-treatment : See below table for ICE classification criteria for mean fluorescein retention

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

UN GHS Classification Combinations of the three ICE Classes
No Category 3×I
2×I, 1×II
1×I, 2×II
No prediction can be made Other combinations

Category 1 3×IV
2×IV, 1×III
2×IV, 1×II*
2×IV, 1×I*
Corneal opacity = 3 at 30 min (in at least 2 eyes)
Corneal opacity = 4 at any time point (in at least 2 eyes)
Severe loosening of epithelium (in at least 1 eye)


Irritation parameter:
percent corneal swelling
Run / experiment:
Experiment 1 & 2 (Physiological saline)
Value:
0
Negative controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
Experiment 1 & 2 (Physiological saline)
Value:
0
Negative controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
Experiment 1 & 2 (Physiological saline)
Value:
0
Negative controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Run / experiment:
Experiment 1 & 2 (Parafilm control)
Value:
0
Negative controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
Experiment 1 & 2 (Parafilm control)
Value:
0
Negative controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
Experiment 1 & 2 (Parafilm control)
Value:
0
Negative controls validity:
valid
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Run / experiment:
Experiment 1 (Distilled TCPD)
Value:
1.1
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Run / experiment:
Experiment 2 (Distilled TCPD)
Value:
1.6
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
Experiment 1 (Distilled TCPD)
Value:
0.5
Remarks on result:
no indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
Experiment 2 (Distilled TCPD)
Value:
0.5
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
Experiment 1 (Distilled TCPD)
Value:
1
Remarks on result:
no indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
Experiment 2 (Distilled TCPD)
Value:
0.83
Remarks on result:
no indication of irritation
Irritation parameter:
percent corneal swelling
Run / experiment:
Experiment 1 (Imidazole)
Value:
24.9
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation parameter:
percent corneal swelling
Run / experiment:
Experiment 2 (Imidazole)
Value:
26.1
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
Experiment 1 (Imidazole)
Value:
4
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation parameter:
cornea opacity score
Run / experiment:
Experiment 2 (Imidazole)
Value:
4
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
Experiment 1 (Imidazole)
Value:
3
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Irritation parameter:
fluorescein retention score
Run / experiment:
Experiment 2 (Imidazole)
Value:
3
Positive controls validity:
valid
Remarks on result:
positive indication of irritation
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: No

DEMONSTRATION OF TECHNICAL PROFICIENCY: The chosen test facility has trained staff technicallly proficient in this OECD 438 test. It is a GLP compliant facility and has adequate historical data to justify the observed results.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Range of historical values if different from the ones specified in the test guideline: As per OECD guidence.

SUMMARY TABLE FOR UN GHS CLASSIFICATION (EXPERIMENT I & 2)

 

Criteria for “No category” (all true)

 

3 endpoints classed as I or 2 endpoints classed as I and

1 endpoint classed as II:

True

No severe corneal morphological changes:

True

Test item was not stuck to the cornea at 240 minutes after the post-treatment rinse:

True

 

Criteria for “Category 1” (one or more true)

 

2 or more endpoints classed as IV:

False

Corneal opacity = 3 at 30 min (in at least 2 eyes):

False

Corneal opacity = 4 at any time point (in at least 2 eyes):

False

Severe loosening of epithelium (in at least 1 eye):

False

 

Criteria for “No prediction can be made” (one or two true)

 

Based on the endpoints not classifiable for No Category, or for Category 1:

False

Particles of test item were stuck to the cornea and could not be washed off during the study:

False


 

Interpretation of results:
GHS criteria not met
Conclusions:
Based on these in vitro eye irritation assays in isolated chicken eyes with Distilled Tricyclopentadiene, the test item was non-irritant, UN GHS Classification: No Category.
Executive summary:

Based on these in vitro eye irritation assays in isolated chicken eyes with Distilled Tricyclopentadiene, the test item was non-irritant, UN GHS Classification: No Category.

Experiment I: No significant swelling (mean ≤5%) was observed during the four-hour observation period on test item treated eyes. No significant cornea opacity change (severity 0.5) was observed on all three eyes. Slight fluorescein retention change (severity 1) was observed on all three eyes. Residual test item was stuck on one cornea surface after the post-treatment rinse, this was cleared at 30 minutes after the post-treatment rinse.

Experiment II: No significant corneal swelling (mean ≤5%) was observed during the four-hour observation period on test item treated eyes. No significant cornea opacity change (severity 0.5) was observed on all three eyes. Slight fluorescein retention change (severity 0.5 on one eye and severity 1 on two eyes) was observed on all three eyes.

All eyes used in the study met the quality control standards. The negative control, parafilm control, and positive control results were within the historical control data range in each experiment. Thus, the study was considered to be valid.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Justification for classification or non-classification

Under the conditions of this study, this substance was not found to be an irritant or corrosive.