4,5-bis(hydroxymethyl)-2-phenyl-1H-imidazole

Endpoint summary

Description of key information

- Skin irritation / corrosion: not a skin irritant (OECD 439, GLP, K, rel.1)

- Eye irritation / damage: no prediction can be made (OECD 438, GLP, K, rel.1)

- Eye irritation / damage: not damaging or irritating (OECD (Q)SAR toolbox v.4.3.0 and Toxtree v3.1.0, K. rel. 2)

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 14 October 2021 to April 2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted according to the OECD TG 439 without any deviation.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

14 June 2021

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature in a dry place
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: Stable under storage conditions
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: NA
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: NA
- Reactivity of the test material with the incubation material used (e.g. plastic ware): NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): NA
- Preliminary purification step (if any): NA
- Final concentration of a dissolved solid, stock liquid or gel: NA
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): NA

FORM AS APPLIED IN THE TEST (if different from that of starting material)
- Specify the relevant form characteristics if different from those in the starting material, such as state of aggregation, shape of particles or particle size distribution: NA

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from multiple donors

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: SkinEthic RHE/ Human Epidermis (RHE/S/17)
- Tissue batch number: 21-RHE-147
- Production date: 21 September 2021 (expiry date: 27 September 2021)
- Shipping date: 21 September 2021
- Delivery date: 21 September 2021
- Date of initiation of testing: 21 September 2021

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

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: 25 x 1mL washes with Phosphate Buffer Saline (DPBS)
- 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: 300 µL
- Incubation time: 3 hours
- Spectrophotometer: ELx800 absorbance microplate reader
- Wavelength: 570 nm
- Filter: n.a.
- Filter bandwidth: n.a.
- Linear OD range of spectrophotometer: Not stated

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: Specification: O.D. > 0.7, Result O.D = 1.1 (CV = 7.1%)
- Barrier function: Specification: 4.0h < ET50 < 10.0h
- Morphology: Multi-layered, highly differentiated epidermis consisting of organised basal, spinous and granular layers, and a multilayered stratum corneum.
- Contamination: Cell have been verified the absence of bacteria, fungus and mycoplasma
- Reproducibility: Not stated

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE: not applicable

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 1

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be irritant to skin if the mean percent viability after 42 minutes exposure and 42 hours (± 1 hour) of post-treatment incubation is > 50%.
- The test substance is considered to be non-corrosive to skin if the mean percent tissue viability after 42 minutes exposure and 42 hours (± 1 hour) of post-treatment incubation is ≤ 50
- Justification for the selection of the cut-off point(s) if different than recommended in TG 439: n/a (same as OECD TG 439)

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 16 mg
- Concentration (if solution): NA

VEHICLE
- Amount(s) applied (volume or weight with unit): NA
- Concentration (if solution): NA
- Lot/batch no. (if required): NA
- Purity:

NEGATIVE CONTROL
- Amount applied (volume) : 16 µL
- Concentration (if solution): NA

POSITIVE CONTROL
- Amount(s) applied (volume): 16 µL
- Concentration (if solution): 5%

Duration of treatment / exposure:

42 minutes

Duration of post-treatment incubation (if applicable):

41 hours ± 1 hour

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1

Value:

123.3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

2

Value:

98.7

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3

Value:

87.3

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: none observed
- Direct-MTT reduction: The direct interaction of MTT with the test item was checked by adding 16 mg of the test item to 300 µL of the solution of MTT at 1 mg/mL (same conditions as in the main test). A yellow solution with white test item was observed after 3 hours of incubation between 37.2°C and 37.4°C, 5% CO2.
Therefore, there is no direct interaction between the test item and MTT.
- Colour interference with MTT: The spectral properties at 570 nm of test item in isopropanol were checked by adding 16 mg of the test item to 1.5 mL of isopropanol (same conditions as in the main test). A white solution was obtained after 2 hours of incubation at ambient temperature with gentle shaking.
The solution was centrifuged for 1 minute at 16000 g before dosing the supernatant.
The mean of the corrected OD after centrifugation was 0.319 which is higher than 0.08 (value corresponding to approximately twice the OD of the extracting solvent).
Therefore, the test item was identified as causing colour interference with the viability assay (mean of the corrected OD > 0.08) and two viable control tissues were added to the study which underwent the entire testing procedure but were incubated with medium instead of MTT solution during the MTT incubation step to generate a non-specific colour (NSC living) control.


DEMONSTRATION OF TECHNICAL PROFICIENCY: yes (cf. 'Attachment')

ACCEPTANCE OF RESULTS: ('cf. Attachment')
- Acceptance criteria met for negative control: yes, OD value of the 3 replicates in the range ≥ 0.8 and ≤ 3.0 and the SD value of the % viability ≤ 18%.
- Acceptance criteria met for positive control: yes, Mean Viability < 40%, and SD value of the % viability ≤ 18%.
- Acceptance criteria met for variability between replicate measurements: OD values of the 3 replicates in the range ≥ 0.8 and ≤ 3.0, and SD value of the % viability is 18.4%.
- Range of historical values if different from the ones specified in the test guideline: cf. attachment.

Table 1: Test item - assessment of the skin irritation individual and average values of OD after 42 minutes exposure

	Well ID	OD	Mean OD/ disc	Mean OD/product	Viability %	Mean viability %	SD	Conclusion
Negative control	SPL_1	0.945 0.921 0.923	0.930	0.868	107.1	100.0	7.9
	SPL_2	0.762 0.778 0.841	0.794		91.4
	SPL_3	0.852 0.896 0.895	0.881		101.5
Positive control	SPL_4	0.011 0.011 0.011	0.011	0.011	1.3	1.3	0.1	Irritant
	SPL_5	0.012 0.012 0.011	0.012		1.4
	SPL_6	0.010 0.013 0.011	0.011		1.3
Test Item PH-21/0755	SPL_7	1.073 1.043 1.096	1.071	0.895	123.3	103.1	18.4
	SPL_8	0.857 0.852 0.861	0.857		98.7
	SPL_9	0.790 0.725 0.759	0.758		87.3
Test Item PH-21/0755 NSC living	SPL_10	0.001 0.001 0.001	0.001	0.001	0.1	0.1	0.1
	SPL_11	0.000 -0.001 -0.001	0.000		0.0
Test item PH-21/-755 Corrected						103.0		Non Irritant

 

Interpretation of results:

GHS criteria not met

Conclusions:

The mean corrected percent viability of the treated tissues was 103.0%. Not classified according to CLP and GHS criteria.

Executive summary:

In an in vitro skin irritation study (Reconstructed Human Epidermis Test Methods - SkinEthic RHE^® model) performed according to the OECD TG No. 439 and in compliance with GLP, 16 mg of undiluted test item was applied to three living reconstructed epidermis for 42 minutes. The application was followed by a rinse with 25 mL of PBS and a 41 hours incubation period at 37°C, 5% CO₂. Cell viability was then measured by enzymatic conversion of the vital dye MTT into a blue formazan salt that was quantitatively measured after extraction from tissues. Additionally, 2 living Human skin model surfaces were treated in the same manner but they were incubated in culture medium instead of MTT solution in order to generate non-specific living color controls.

 

The mean corrected percent viability of the treated tissues was 103.0% for the test item and 1.3% for the positive control (5% SDS).

The quality criteria required for acceptance of results in the tests were satified:

• SD ≤ 18%


• Negative control: OD value of the 3 replicates in the range ≥ 0.8 and ≤ 3.0 and the SD value of the % viability ≤ 18%.


• Positive control: Mean Viability < 40%, and SD value of the % viability ≤ 18%.

 

In accordance with the Regulation EC No. 1272/2008, the test item CUREZOL 2PHZ-PW is not considered to be irritating to the skin and therefore classified as a Non-irritant UN GHS 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:

weight of evidence

Study period:

From 20 August 2021 to April 2022

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study conducted according to the OECD TG 438 with minor deviations not affecting the reliability of the study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

25 June 2018

Deviations:

yes

Remarks:

During preparation, eye were dissected from the skull before the assessment of corneal integrity. This deviation has no impact on the study reliability as it has been validated by there CRO and is part of their standard SOP.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Keep away from heat and sources of ignition. Store in tightly closed container. Store in a well-ventilated place. Store in a cool, dark place at room temperature (20°C ± 10°C)
- Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: NA
- Stability in the medium, i.e. sensitivity of the test material to hydrolysis and/or photolysis: NA
- Solubility and stability of the test material in the solvent/vehicle and the exposure medium: NA
- Reactivity of the test material with the incubation material used (e.g. plastic ware): NA

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing (e.g. warming, grinding): NA
- Preliminary purification step (if any): NA
- Final concentration of a dissolved solid, stock liquid or gel: NA
- Final preparation of a solid (e.g. stock crystals ground to fine powder using a mortar and pestle): NA

FORM AS APPLIED IN THE TEST (if different from that of starting material)
- Specify the relevant form characteristics if different from those in the starting material, such as state of aggregation, shape of particles or particle size distribution: NA

Species:

chicken

Strain:

not specified

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: Etablissement Brun, 33820 Etauliers, France
- Number of animals: Not stated
- Characteristics of donor animals (e.g. age, sex, weight):Spring chickens, 7 weeks old, 1.5 - 2.5kg
- Storage, temperature and transport conditions of ocular tissue (e.g. transport time, transport media and temperature, and other conditions): The intact heads were transported from the slaughterhouse at ambient temperature in plastic boxes humidified with towels moistened with physiological saline. Heads collected from source at 8:45am and eyes enucleated at ICARE at 10.53am.
- Time interval prior to initiating testing: 45 minutes
- Indication of any existing defects or lesions in ocular tissue samples: Eyes where ocular lesions were present were discarded prior to testing.
- Indication of any antibiotics used: Not stated
- Selection and preparation of corneas: Not stated
- Quality check of the isolated corneas: Examination under slit-lamp microscope (Haag-Streit BP900 with a slit width of 0.095mm)

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied (weight): 30 mg
- Concentration (if solution): NA

VEHICLE
- Amount(s) applied (volume or weight with unit): NA
- Concentration (if solution): NA
- Lot/batch no. (if required): NA
- Purity: NA

Duration of treatment / exposure:

10 second exposure

Duration of post- treatment incubation (in vitro):

30, 75, 120, 180 and 240 minutes

Number of animals or in vitro replicates:

3

Details on study design:

SELECTION AND PREPARATION OF ISOLATED EYES: The selection of eyes was based on the cornea integrity following excision.

NUMBER OF REPLICATES: 3 replicates

NEGATIVE CONTROL USED: Physiological saline – Dutscher Batch No. C0543A01 – CAS : 7647-14-5 – EC: 231-598-3 – SMILES: [Na+].[Cl-] – Storage : room temperature

POSITIVE CONTROL USED: sodium hydroxide – Fisher Scientific, Batch No. 0000080257 - CAS : 1310-73-2 – EC: 215-185-5 – SMILES: [OH-].[Na+] – Storage : room temperature

APPLICATION DOSE AND EXPOSURE TIME: 30 mg of test material was used with 10s exposure time. XX was used for both control substances with 10 s exposure time

OBSERVATION PERIOD: NA

REMOVAL OF TEST SUBSTANCE
- Volume and washing procedure after exposure period: Two rinses with 10mL physiological saline followed by the use of a cotton bud soaked in saline to remove residual test material
- Indicate any deviation from test procedure in the Guideline: Use of cotton bud soaked with saline. However justification in the form of validated test methods does not deem this a deviation that affects the integrity of the test.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Haag-Streit BP900 slit-lamp microscope with depth-measuring device no. I. For the measurement of corneal thickness, the slit-width was set at 9½, equalling 0.095 mm
- Damage to epithelium based on fluorescein retention: NA
- Swelling: measured with optical pachymeter on a slit-lamp microscope; slit-width setting: Haag-Streit BP900 slit-lamp microscope with depth-measuring device no. I. For the measurement of corneal thickness, the slit-width was set at 9½, equalling 0.095 mm
- Macroscopic morphological damage to the surface: Not stated
- Others (e.g, histopathology): NA

SCORING SYSTEM: as per OECD TG 438

DECISION CRITERIA: as per OECD TG 438

Irritation parameter:

cornea opacity score

Run / experiment:

45 minutes pretreatment, 30, 75, 120, 180 and 240 minutes post treatment

Value:

0

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Remarks:

ICE class I

Irritation parameter:

fluorescein retention score

Run / experiment:

45 minutes pre-test and 30 minutes post treatment

Value:

>= 0.5 - <= 2

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Remarks:

ICE Class III

Irritation parameter:

cornea opacity score

Run / experiment:

45 minutes pre test, 30, 76, 120, 180 and 240 minutes post treatment

Value:

>= 0.5 - <= 0.62

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Remarks:

ICE Class II

Irritation parameter:

percent corneal swelling

Run / experiment:

45 minutes pre test, 30, 76, 120, 180 and 240 minutes post treatment

Value:

>= 3 - <= 9

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Remarks:

ICE Class II

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: none observed

DEMONSTRATION OF TECHNICAL PROFICIENCY: cf 'Attachment"

ACCEPTANCE OF RESULTS: cf. "'Attachments'
- Acceptance criteria met for negative control: The combination of the three endpoints for the negative control, physiological saline, was 3 x I. Therefore, as expected the negative control is classified as “No Category”, as expected.
- Acceptance criteria met for positive control: The combination of the three endpoints for the positive control, Sodium Hydroxide 5%, was
3 x IV. Therefore, as predicted the positive control is classified as “Corrosive/Severe Irritant.
- Range of historical values if different from the ones specified in the test guideline: as per OECD TG 438. Cf. 'Attachment'

Table 2: Negative control - Assessment of the eye corrosion/severe irritation individual and average values for evaluation of corneal lesions after treatment

Endpoint measured	Eye no.	Time (min)
		-45	30	75	120	180	240
Corneal opacity	16	0	0	0	0	0	0
Mean		0.0	0.0	0.0	0.0	0.0	0.0
ICE class			I	-	-	-	-
Fluorescein retention	16	0.5	0.5	-	-	-	-
Mean		0.5	0.5
ICE class		I
Corneal thickness	16	0.51	0.51	0.51	0.52	0.52	0.52
Corneal swelling (%)	16	-	0	0	2	2	2
Mean		-	0	0	2	2	2
ICE class		I
Combination of the 3 endpoints		3 x I
Classification		No Category

Table 3: Positive control - Assessment of the eye corrosion/severe irritation individual and average values for evaluation of corneal lesions after treatment

Endpoint measured	Eye no.	Time (min)
		-45	30	75	120	180	240
Corneal opacity	1	0	4	4	4	4	4
	2	0	4	4	4	4	4
	3	0	4	4	4	4	4
Mean		0.0	4.0	4.0	4.0	4.0	4.0
ICE class		IV
Fluorescein retention	1	0.5	3	-	-	-	-
	2	0.5	3	-	-	-	-
	3	0.5	3	-	-	-	-
Mean		0.5	3	-	-	-	-
ICE class		IV
Corneal thickness	1	0.54	0.84	0.98	1.05	1.10	1.11
	2	0.53	0.81	0.86	0.91	1.05	1.05
	3	0.56	0.82	0.87	0.89	1.03	1.12
Corneal swelling (%)	1	-	56	81	94	104	106
	2	-	53	62	72	96	98
	3	-	46	55	59	84	100
Mean		-	52	66	75	95	101
ICE class		IV
Combination of the 3 endpoints		3 x IV
Classification		Category I: ‘Corrosive/Severe Irritant’

Table 4: Test item Curezol 2PHZ-PW - Assessment of the eye corrosion/severe irritation individual and average values for evaluation of corneal lesions after treatment

Endpoint measured	Eye no.	Time (min)
		-45	30	75	120	180	240
Corneal opacity	7	0	4	0	0	0	0
	8	0	4	0	0	0	0
	9	0	4	0	0	0	0
Mean		0.0	0.0	4.0	0.0	0.0	0.0
ICE class		I
Fluorescein retention	7	0.5	2	-	-	-	-
	8	0.5	2	-	-	-	-
	9	0.5	2	-	-	-	-
Mean		0.5	2	-	-	-	-
ICE class		III
Corneal thickness	7	0.58	0.59	0.59	0.59	0.62	0.62
	8	0.50	0.53	0.53	0.53	0.57	0.59
	9	0.54	0.54	0.54	0.54	0.54	0.55
Corneal swelling (%)	7	-	2	2	2	7	7
	8	-	6	6	6	14	18
	9	-	0	0	0	0	2
Mean		-	3	3	3	7	9
ICE class		III
Combination of the 3 endpoints		1 x I 1 x II 1 x III
Classification		No prediction can be made

Interpretation of results:

study cannot be used for classification

Conclusions:

The combination of the three endpoints for the test item CUREZOL 2PHZ-PW was 1 x I, 1 x II, 1 x III. No prediction can be made.

Executive summary:

In an in vitro eye irritation / corrosion study (Isolated Chicken Eye) performed according to the OECD TG No. 438 and in compliance with GLP, 30 mg of undiluted test item was applied to three enucleated chicken eyes for 10 seconds. The eyes were then rinsed twice with 10 mL of physiological saline. Following two rinses remnants of the product was still evident on the entire cornea of all three eyes. The remaining test substance was removed using a cotton bud soaked with physiological saline soaked. Three eyes were treated in the same manner with a positive control and one eye with a negative control. Damages by the test item were assessed by determination of corneal swelling, opacity, and fluorescein retention 45 minutes prior the treatment and at 30, 75, 120, 180 and 240 minutes  (± 5) minutes post treatment rinse.

 

The ocular reactions observed in eyes treated with the test item were:

• maximal mean score of corneal opacity: 0.0, corresponding to ICE class I;


• mean score of fluorescein retention: 2.0, corresponding to ICE class III;


• maximal mean corneal swelling: 9 %, corresponding to ICE class II.

 

The combination of the three endpoints for the test item CUREZOL 2PHZ-PW was 1 x I, 1 x II, 1 x III.

The combination of the three endpoints for the positive control, Sodium Hydroxide 5%, was 3 x IV. Therefore, the positive control is classified as “Corrosive/Severe Irritant”, as expected.

The combination of the three endpoints for the negative control, physiological saline, was 3 x I. Therefore, the negative control is classified as “No Category”, as expected.

 

In accordance with the Regulation (EC) No. 1272/2008, the results obtained under these experimental conditions lead to the category “no prediction can be made”, as defined by the OECD guideline No.438. Therefore, on prediction can be made on the test item CUREZOL 2PHZ-PW and additional testing is required to establish a definitive classification.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

TBC (eye)

 

Skin irritation / corrosion:

A study is available (ICARE, 2022).This in vitro skin irritation study (Reconstructed Human Epidermis Test Methods - SkinEthic RHE^® model) was performed according to the OECD TG No. 439 and in compliance with GLP.

The mean corrected percent viability of the treated tissues was 103.0% for the test item and 1.3% for the positive control (5% SDS).

The quality criteria required for acceptance of results in the tests were all satified.

The test item CUREZOL 2PHZ-PW is not considered to be irritating to the skin under the test conditions.

 

Eye irritation / damage:

A study is available (ICARE, 2022). This in vitro skin irritation / corrosion study (Isolated Chicken Eye Test) was performed according to the OECD TG No. 438 and in compliance with GLP.

The ocular reactions observed in eyes treated with the test item were:

• maximal mean score of corneal opacity: 0.0, corresponding to ICE class I;


• mean score of fluorescein retention: 2.0, corresponding to ICE class III;


• maximal mean corneal swelling: 9 %, corresponding to ICE class II.

The combination of the three endpoints for the test item CUREZOL 2PHZ-PW based on class was:

1 x I;

1 x II;

1 x III.

The combination of the three endpoints for the positive control, Sodium Hydroxide 5%, was 3 x IV. Therefore, the positive control is classified as “Corrosive/Severe Irritant”, as expected.

The combination of the three endpoints for the negative control, physiological saline, was 3 x I. Therefore, the negative control is classified as “No Category”, as expected.

The results obtained under these experimental conditions for the test item lead to the category “no prediction can be made”, as defined by the OECD guideline No.438. This due to the presence of 3 categories below category IV, but one catgory at level III.  

 

In line with ECHA R.7a, ECHA R.6 and ECHA Guidance on the Application of the CLP Criteria, Figure 3.4 (pg 307: v5.0, 2017),  in order to build a weight of evidence for the eye irritation/damage endpoint requird at Annex VII (Section 8.2) predictions were made using the OECD QSAR toolbox v4.3 and Toxtree v3.1.0 and the data on skin corrosion/irritation was used. All results are considered in relation to corresponding information presented and in accordance with the tonnage driven information requirements of REACH Regulation (EC) 1907/2006 in a weight of evidence. Specifically:

1. No prediction could be made following assessment with the Isolated Chicken Eye (ICE) Assay (OECD TG 438);


2. The test substance was not classified as an irritant to the skin following In Vitro Skin Irritation study ( OECD TG 439);


3. The test substance was not predicted to be irritating or corrosive in OECD QSAR toolbox v4.3. The substance was within the exclusion rules indicated by the BfR – Group all melting point > 200 C (This exclusion rule has been found to be 99-100 % accurate (see QMRF)); No inlcusion rules were met for Toxtree v3.1.0 or OECD QSAR toolbox v4.3.  No exclusion rules were met in Toxtree v3.1.0 but these exclusions are out of date (2008) as indicated by the OECD QSAR toolbox v4.3 (2010) 


4. The water solubility is considered to be: 0.194 g/L ( OECD 105)


5. A melting point could not be determined as the test substance degrades prior to melting: Not determined (OECDTG 102)


6. The log K_OW is considered to be: 1.45 (OECD TG 117)

In conclusion, with the presence of exclusion criteria and no inclusion criteria during the execution of a valid (Q)SAR model, taken in combination with the weight of evidence from experimental data on skin irritation/corrosion (not irritating or corrosive) and the substance's physico-chemical properties, the substance is concluded as having no damaging or irritating effects to the eye.The weight of evidence indicates the substance should not be classified for eye irritation or corrosion under Regulation (EC) 1272/2008 as amended.

Justification for classification or non-classification

Harmonised classification:

The substance has no harmonised classification according to the Regulation (EC) No. 1272/2008 (CLP).

 

Self classification:

Based on the available information no additional self-classification is proposed regarding both skin and eye irritation according to the CLP and to the GHS.

No data was available regarding respiratory irritation, however the substance not being classified for skin and eye irritation, no classification is expected for respiratory irritation.