Sodium bis[3-[[1-(3-chlorophenyl)-4,5-dihydro-3-methyl-5-oxo-1H-pyrazol-4-yl]azo]-4-hydroxy-N-methylbenzenesulphonamidato(2-)]cobaltate(1-)

Endpoint summary

Description of key information

The test item did not cause corrosive effects in an in vitro assay (BCOP) nor did it cause significant irritation in rabbit eyes in vivo.

The test item did not cause irritant effects in an in vitro human skin model assay.

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:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: according to OECD 439 Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: UN GHS (published 2003, last (3rd) revision 2009

Deviations:

no

Principles of method if other than guideline:

Based on a “Statement on the Scientific Validity of In Vitro Tests for Skin Irritation” of the European Commission (November 2008), official acceptance of the test method in the EU was achieved and implemented in EU, 2008a, Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to EC Regulation No 1907/2006 of the European Parliament and of the Council on REACH; 1st ATP 2009: EC Regulation No 761/2009 of 23 July 2009 amending, for the purpose of its ATP, EC Regulation No 440/2008 laying down test methods pursuant to EC Regulation No 1907/2006 of the European Parliament and of the Council on REACH, section B46.

GLP compliance:

yes (incl. QA statement)

Species:

other: reconstituted human epidermis model

Strain:

other: reconstituted human epidermis model

Type of coverage:

other: Topical

Preparation of test site:

other: Not applicable

Vehicle:

other: No vehicle used

Amount / concentration applied:

Test Item Preparation:
The test material was crushed and ground in a mortar with pestle to improve the consistency. Each approximately 25 mg (~ 39 mg/cm2 according to guideline) of the test item were applied to the tissues, wetted with 25 µL DPBS, and spread to match the surface of the tissue for a complete treatment time of 60 minutes.

Negative Control:
30 µL DPBS (MatTek) was used as negative control per tissue.

Positive Control:
30 µL of a 5% SLS solution in deionised water (MatTek) was used a positive control per tissue, freshly prepared prior to the start of the experiment.


Each three tissues of the human skin model EpiDerm™ were treated with the test item, the negative or the positive control for 60 minutes.

Approximately 25 mg of the test item were applied to each tissue, wetted with 25 µL of DPBS, and spread to match the surface of triplicate tissue.
30 µL of either the negative control (DPBS) or the positive control (5% SLS) were applied to triplicate tissue each.

The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 43 hours the tissues were treated with the MTT solution for 3 hours following 69.75 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

Duration of treatment / exposure:

60 minutes

Observation period:

Not applicable

Number of animals:

Not applicable

Details on study design:

Test for Direct MTT Reduction and Colour Interference
For correct interpretation of results it was necessary to assess the ability of the test item to directly reduce MTT. For this purpose, approximately 25 mg of the test item were added to 1 mL of MTT solution (MTT (Sigma, Germany) concentrate diluted with MTT diluent (DMEM, Gibco, Germany) (resulting: 1 mg/mL)). This mixture was incubated in the dark at 37 ± 1.5 °C (5 ± 0.5% CO2) for 60 minutes.
Since the colour did not turn blue/purple, the test item was not considered to be a MTT reducer.
Prior to the start of the test, the test item’s colour interference potential had to be evaluated. For this purpose approximately 25 mg of the test item were mixed with 300 µL of deionised water. This mixture was incubated for 60 minutes at 37 ± 1.5 °C (5 ± 0.5% CO2).
Since the colour of the test item / water mixture was intensive, an additional functional check was performed. The coloured test item was applied to one viable tissue, which underwent the entire skin irritation test (exposure period: 60 minutes; recovery period: about 41 hours; extraction period: 70 hours), but was incubated with medium instead of MTT solution during the MTT incubation step.
Since the OD of the tissue treated by the coloured test item was > 30% (651.8%) of the DPBS treated control tissue, the test item could have been considered as incompatible with the test. But according to an expert judgment test item particles could have been trapped in the tissue and could not be removed in the washing step. The gained high absorption value was most probably caused by reflection of the remaining particles.
Therefore, the real MTT OD (unaffected by interference with the coloured test items) was calculated using following formula:
OD = ODcoloured tissue (MTT assay) – ODcoloured tissue (no MTT assay)

Performance
Pre-warming of EpiDerm™ Tissues
One day prior to the performance, the plastic bag containing the 24-well plate with epidermal tissues was opened under sterile conditions. Under an airflow using forceps, the gauze was removed and the inserts were taken out. Any remaining agarose that adheres to the outer sides of the inserts was removed by gentle blotting on the sterile filter paper or gauze, and the tissues were placed in the empty, sterile 6-well plate. Prior to the exposure of the test item and of the controls the EpiDerm™ tissues were inspected for quality: If necessary, it was taken care, that
1. air bubbles between agarose and insert were not > 30% of the total surface,
2. liquid on top of the insert was removed with steriles cotton tips,
3. if again moisture is observed on top of the inserts after the pre-incubation or in
case of visible defects the respective skin models were discarded.
0.9 mL of the assay medium (20 – 25 °C) was pipetted into each well of sterile 6-well plates. The inserts with the EpiDerm™ tissues were placed in the upper wells, and were pre-incubated for 60 minutes in the incubator (37 ± 1.5 °C, 5 ± 1% CO2, 95 ± 5% RH). Following, the inserts were transferred from upper wells into the lower wells of the 6-well plates, and, the pre-incubation was continued for further 22 hours and 20 minutes (37 ± 1.5 °C, 5 ± 1% CO2, 95 ± 5% RH).
Treatment
After pre-incubation of EpiDerm™ tissues was completed, medium was replaced by 0.9 mL of fresh medium per well. The negative and positive control, the vehicle control, and the test item were added into the insert atop the corresponding EpiDerm™ triplicate tissues. The treatment time was 60 minutes in total. Within this period the 6-well plates were put into the incubator for 35 minutes at 37 ± 1.5 °C, 5 ± 0.5 % CO2. In the remaining period the plates were placed in a sterile bench at room temperature until the end of treatment.
After the end of the treatment interval the inserts were removed immediately from the 6-well plate and tissues were gently rinsed with DPBS at least 15 times in order to remove any residual test material. After the rinsing the inserts were submerged in DPBS at least three times. Afterwards the inserts were once again rinsed with sterile DPBS from the inside and the outside. Excess DPBS was removed by gently shaking the inserts and blotting the bottom with sterile blotting paper. The tissues were carefully dried using sterile cotton tipped swap. The tissues were then transferred into new 6-well plates with 0.9 mL of fresh assay medium in the upper row. The inserts were placed in the prepared holding plate. Tissues were incubated for 23 hours and 38 minutes at 37 ± 1.5 °C, 5 ± 0.5 % CO2. After incubation the inserts were transferred into new 6-wells plates containing fresh medium. Thereafter tissues were incubated for another 19 hours at 37 ± 1.5 °C, 5 ± 0.5 % CO2. The complete incubation time was 42 hours and 30 minutes.

MTT Assay
On the day of testing the MTT concentrate was diluted with the MTT diluent (1 mg/mL). The 24-well plates were prepared before the end of the tissue pre-warming period. A volume of 300 µL of the MTT solution was added to each well and the plates were kept in an incubator (37 ± 1 °C, 5 ± 0.5 % CO2) until further use.
After the 42-hours incubation period was completed for all tissues and exposure groups, culture inserts were transferred from the holding plates to the MTT-plates. After a 3-hour incubation period (37 ± 1 °C, 5 ± 0.5 % CO2), the MTT solution was aspirated from the wells, and the wells were rinsed three times with DPBS. Inserts were transferred onto new 24-well plates. The inserts were immersed into extractant solution by gently pipetting 2 mL extractant solution (isopropanol) in each insert. The level rose above the upper edge of the insert, thus tissues were completely covered from both sides. The 24-well plate was sealed to inhibit the isopropanol evaporation.
The formazan salt was extracted for 70 hours without shaking in the refrigerator.
After the extraction period was completed, the inserts were pierced with an injection needle to allow the extract to run into the well from which the insert was taken. Afterwards the insert was discarded. The 24-well plates were placed on a shaker for 15 minutes until the solution was homogeneous in colour.
Per each tissue, 3  200 µL aliquots of the blue formazan solution were transferred into a 96-well flat bottom microtiter plate from the 15 minutes exposure. OD was read in a microplate reader (Versamax® Molecular Devices, Softmax Pro, version 4.7.1) with a 570 ± 1 nm filter. Mean values were calculated from the 3 wells per tissue.

Irritation / corrosion parameter:

% tissue viability

Value:

>= 84.5

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

threshold for irritancy:≤50%

Irritant / corrosive response data:

Example:
Compared to the relative absorbance value of the negative control the mean relative absorbance value was reduced to 84.5% (corrected value) after exposure of the skin tissues to the test item. This value is above the threshold for irritancy of ≤ 50%. Therefore, the test item is not considered to possess an irritant potential.

Other effects:

No

Results after treatment with the test item, and the controls

Dose Group	Treat-ment Interval	Absor-bance 570 nm Tissue 1*	Absor-bance 570 nm Tissue 2*			Absor-bance 570 nm Tissue 3*		Mean Absor-bance of 3 Tissues			Rel. Absor-bance [%] Tissue 1, 2 + 3**		Relative Standard Deviation [%]	Mean Rel. Absorbance [% of Negative Control]***
Negative Control	60 min	1.869	1.894			1.949		1.904			98.2 99.5 102.3		2.1	100.0
Positive Control	60 min	0.102	0.105			0.074		0.094			5.4 5.5 3.9		18.0	4.9
Test Item	60 min	1.578	1.582			1.713		1.624			82.9 83.1 90.0		4.7	85.3
Colour interference pre-experiment: Functional Test without MTT
		Absorbance 570 nm Well 1			Absorbance 570 nm Well 2		Absorbance 570 nm Well 3			Mean Absorbance of 3 Wells		Mean Absorbance subtracted by blank		Mean Rel. Absorbance [% of Negative Control]***
Negative Control	60 min	0.0381			0.04		0.0398			0.0393		0.0028		100.0
Test Item without MTT	60 min	0.0617			0.0527		0.0504			0.0549		0.0185		651.8
Data correction procedure
	Mean Absorbance of 3 Tissues (main experiment)			Mean Absorbance of 3 Wells (Functional Test without MTT)					ODMTT– ODno MTT			Corrected Mean Rel. Absorbance [% of Negative Control]***
Negative Control	1.904			0.0028					1.901			100.0
Test Item	1.624			0.0185					1.606			84.5

* Mean of three replicate wells after blank correction

**  Relative absorbance per tissue [rounded valuea]: 100 x absorbance_^tissue

(mean absorbance _{negative control})

*** Relative absorbance per treatment group [rounded values]:  100 x (mean absorbance _{test item/positive control})

(mean absorbance _{negative control})

Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour. Due to the intrinsic colour of the test item, an additional test with viable tissues (entire test procedure) but without MTT was performed. A correction from the mean relative absorbance value of the test item, corresponding to the cell viability was necessary.

After correction of absorbance (OD = absorbance_MTT– absorbance_{no MTT}) the mean relative absorbance of Savinyl-Gelb 2RLS, trocken, corresponding to the cell viability was 84.5% (threshold for irritancy:≤50%), consequently the test item was not irritant to skin.

 

 

 

 

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item is not irritant to skin according to UN GHS and EU CLP regulation.

Executive summary:

This in vitro study was performed to assess the irritation potential by means of the Human Skin Model Test.

The test item passed the MTT- pre-test. Due to its intrinsic colour, an additional test with one viable tissue (entire test procedure), but with medium instead of MTT addition was performed.

Since the OD of the tissue treated by the coloured test item was > 30% (651.8%) of the DPBS treated control tissue in this test,the test item could have been considered as incompatible with the test. But according to anexpert judgment test item particles were most probably trapped in the tissue and could not be removed in the washing step. The gained high absorption value was most probably caused by reflection of the remaining particles.

Therefore, the real MTT OD (unaffected by interference with the coloured test items) was calculated by subtracting the OD_{no MTT}from the OD_{with MTT.}

The test item, the negative control (DPBS), and the positive control (5% SLS) were applied to each of triplicate tissue.

The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 41 hours and 50 minutes the tissues were treated with the MTT solution for 3 hours following 70 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD at or above 0.8 and ≤ 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.

Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 4.9% thus ensuring the validity of the test system.

The relative standard deviations between the % variabilities of the test item, the positive and negative controls in the main test were 18% (threshold of the "OECD Guideline for the Testing of Chemicals 439 :In vitro Skin Irritation: Reconstructed Human Epidermis Test Method”: ≤ 18%), thus ensuring the validity of the study.

Compared to the relative absorbance value of the negative control the corrected mean relative absorbance value was reduced to 84.5% (corrected value) after exposure of the skin tissues to the test item. This value is above the threshold for irritancy of ≤ 50%. Therefore, the test item is not considered to possess an irritant potential.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), Testing Guidelines for Toxicology Studies, 12 NohSan No. 8147

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Two New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Envigo RMS (UK) Limited, Leicestershire, UK. At the start of the study the animals weighed 2.80 or 3.06 kg and were 12 to 20 weeks old. After an acclimatization period of at least 5 days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Envigo RMS (UK) Limited, Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give 12 hours continuous light (06:00 to 18:00) and 12 hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Vehicle:

unchanged (no vehicle)

Controls:

other: The left eye remained untreated and was used for control purposes.

Amount / concentration applied:

0.1 mL of the test item, which was found to weigh approximately 57 mg

Duration of treatment / exposure:

up to 1 hour

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

2

Details on study design:

Immediately before the start of the test, both eyes of the provisionally selected test rabbits were examined for evidence of ocular irritation or defect with the aid of a light source from a standard ophthalmoscope. Only animals free of ocular damage were used.

Initially, a single rabbit was treated. A subcutaneous injection of buprenorphine 0.01 mg/kg was administered 60 minutes prior to test item application to provide a therapeutic level of systemic analgesia. Five minutes prior to test item application, a pre dose anesthesia of ocular anesthetic (two drops of 0.5% tetracaine hydrochloride) was applied to each eye.

A volume of 0.1 mL of the test item, which was found to weigh approximately 57 mg (as measured by gently compacting the required volume into an adapted syringe) was placed into the conjunctival sac of the right eye, formed by gently pulling the lower lid away from the eyeball. The upper and lower eyelids were held together for about one second immediately after treatment, to prevent loss of the test item, and then released. The left eye remained untreated and was used for control purposes. Immediately after administration of the test item, an assessment of the initial pain reaction was made according to the six point scale shown in Appendix 1.

Eight hours after test item application, a subcutaneous injection of post dose analgesia, buprenorphine 0.01 mg/kg and meloxicam 0.5 mg/kg, was administered to provide a continued therapeutic level of systemic analgesia. The treated animal was checked for signs of pain and suffering approximately 12 hours later. No further analgesia was required.

After consideration of the ocular responses produced in the first treated animal, a second animal was similarly treated.

Assessment of ocular damage/irritation was made approximately 1 hour and 24, 48 and 72 hours following treatment, according to the numerical evaluation (Draize, J.H, 1977) given in Appendix 2.

Any other ocular effects were also noted. Examination of the eye was facilitated by the use of the light source from a standard ophthalmoscope.

Any clinical signs of toxicity, if present, were also recorded.

Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Irritation parameter:

cornea opacity score

Basis:

animal: 75164 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

other: No effects observed

Irritation parameter:

cornea opacity score

Basis:

animal: 75174 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

4

Reversibility:

not specified

Remarks:

No effects observed

Irritation parameter:

iris score

Basis:

animal: 75164 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Irritation parameter:

iris score

Basis:

animal: 75174 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0

Max. score:

2

Reversibility:

other: No effects observed

Irritation parameter:

other: redness

Basis:

animal: 75164 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

other: redness

Basis:

animal: 75174 Male

Time point:

other: Mean 24, 48 and 72 hours

Score:

0.7

Max. score:

3

Reversibility:

fully reversible within: 72 hours

Irritation parameter:

chemosis score

Basis:

animal: 75164 Male

Time point:

other: Mean 24, 48 and 72

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritation parameter:

chemosis score

Basis:

animal: 75174 Male

Time point:

other: 24, 48 and 72 hours

Score:

0.3

Max. score:

4

Reversibility:

fully reversible within: 48 hours

Irritant / corrosive response data:

Individual and mean scores for ocular irritation are given in Table 1.
Red/brown colored staining of the fur around the treated eye was noted in both animals at all observations.
No corneal or iridial effects were noted during the study.
Moderate conjunctival irritation was noted in both treated eyes 1 hour after treatment with minimal conjunctival irritation noted at the 24 and 48 Hour observations.
Both treated eyes appeared normal at the 72Hour observation.
No corrosive effects were noted during the study. The test item did not induce significant or irreversible damage to the rabbit eye.

Other effects:

Body weight
Individual body weights and body weight change are given in Table 2.
One animal showed body weight loss and the other animal showed expected gain in body weight during the study.

Interpretation of Results

Data were summarized in tabular form, showing for each animal the irritation scores for the designated observation time, a description of the degree and nature of irritation, the presence of serious lesions and non‑ocular effects. The scores of each animal at the following reading times (24, 48 and 72 hours) were used in calculating the respective mean values for each type of lesion.

 

The results were interpreted according to Regulation (EC) No 1272/2008 of the European Parliament and of the Council of 16 December 2008, relating to the Classification, Labelling and Packaging of Dangerous Substances.

Table 1     Eye Irritation Scores

Individual Scores for Eye Irritation

Rabbit Number and Sex	IPR	Evaluation interval	Corneal Opacity	Area of Corneal Opacity	Iris	Conjunctivae
						Redness	Chemosis	Discharge
75164Male	0	1 Hour	0	0	0	2	2	1Sf
75174Male	0		0	0	0	2	1	1Sf
75164Male		24 Hour	0	0	0	1	1	0Sf
75174Male			0	0	0	1	1	0Sf
75164Male		48 Hour	0	0	0	1	0	0Sf
75174Male			0	0	0	1	0	0Sf
75164Male		72 Hour	0	0	0	0	0	0Sf
75174Male			0	0	0	0	0	0Sf

IPR=Initial pain reaction

Sf =       Red/brown colored staining of the fur around the treated eye

Table 1 (continued)  Eye Irritation Scores

Mean Values after 24, 48 and 72 Hours

Rabbit Number and Sex	Number of available data points	Corneal Opacity	Iris	Conjunctivae
				Redness	Chemosis
75164Male	3	0.0	0.0	0.7	0.3
75174Male	3	0.0	0.0	0.7	0.3

 

Assessment According to Regulation (EC) No. 1272/2008

Evaluated Intervals	Corneal Opacity	Iris	Conjunctivae
			Redness	Chemosis
24 Hours	Not classified	Not classified	Not classified	Not classified
48 Hours
72 Hours

 

Table 2     Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 3
75164Male	2.80	2.75	-0.05
75174Male	3.06	3.10	0.04

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to the findings in this study, the test item does not meet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

Executive summary:

The primary eye irritation potential of the test item was investigated according to a method compatible with OECD test guideline No. 405 and Method B5. The test item was applied by instillation of 0.1 mL into the right eye of each of two young adult New Zealand White rabbits. Scoring of irritation effects was performed approximately 1, 24, 48 and 72 hours after test item instillation. 

 

The mean score was calculated separately for each animal across three scoring times (24, 48 and 72 hours after instillation) for corneal opacity, iritis, redness and chemosis of the conjunctivae. The individual mean scores for corneal opacity and iritis was 0.0 for both animals. The individual mean scores for the conjunctivae were 0.7 for reddening and 0.3 for chemosis. 

 

The instillation of the test item into the eye resulted in conjunctival irritation. These effects were reversible and were no longer evident 72 hours after treatment in both animals (end of the observation period). No abnormal findings were observed in the cornea of any animal at any of the examinations. No corrosion was observed at any of the measuring intervals. No clinical signs of toxicity were observed.

 

Thus, the test item did not induce significant or irreversible damage to the rabbit eye.

 

According to the findings in this study, the test item does notmeet the criteria for classification according to Regulation (EC) No. 1272/2008 of the European Parliament and of the Council of 16 December 2008.

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

The test item was not classified for eye irritation, as in vitro as well as in vivo assays did not show relevant irritation potential.

The test item was not classified for skin irritation, as in vitro human skin model assays did not show relevant irritation potential.