Sodium 2-butoxyethyl sulphate

Endpoint summary

Description of key information

Skin irritation (OECD 431), EPISKIN: not corrosive

Skin irritation (OECD 439), EPISKIN: not irritative

Eye irritation (OECD 438), chicken eyes: corrosive

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

10 Oct 2017 - 21 Nov 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

29 July 2016

GLP compliance:

yes (incl. QA statement)

Remarks:

Országos Gyógyszerészeti és Élelmezés-egészégügyi Intézet

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Adult donors

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: reconstituted three-dimensional human skin model EPISKIN-SM™ (SkinEthic)
- Tissue batch number(s): 17-EKIN-041
- Expiration date: 16 October 2017

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37°C
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: EpiSkinTMSM units were rinsed thoroughly with approximately 25 mL PBS 1x solution to remove the test item from the epidermal surface.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 2 mL MTT 0.3 mg/mL solution
- Incubation time: 3 h
- Spectrophotometer: Yes, plate spectrophotometer
- Wavelength: 570 nm ± 10nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Barrier function: Specification 1.5 mg/mL ≤ IC50 ≥ 3.0 mg/mL; result: 1.9 mg/mL
- Morphology: Specification ≥ 19.5; result: 23.4±0.4, CV = 1.6%
- Contamination: Screened for HIV-1, hepatits B and C, bacteria, yeast, and other fungi. No contaminates were detected.

NUMBER OF REPLICATE TISSUES: In this assay 2 replicates of test item, 2 replicates negative control and 2 replicates positive control were used.

PREDICTION MODEL / DECISION CRITERIA
A substance was considered corrosive Category 1A if tissue viabilities were < 35% after 3 min exposure. A substance was considered corrosive Category 1B/1C if tissue viabilities were ≥ 35% after 3 min exposure AND < 35% after 1 h exposure OR ≥ 35% after 1 hr exposure AND < 35% after 4 h exposure. A substance was considered noncorrosive if tissue viabitilites were ≥ 35% after 4 h exposure.

The test meets acceptance criteria if:
- The mean OD value of the two negative control tissues was 0.965.
- The positive control result showed 2% viability.
- The difference of viability between the two tissue replicates: Negative control: 15%; Positive control: 1%; Test item: 28%

Control samples:

yes, concurrent negative control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 20 mg test item was added to 2 mL MTT 0.3 mg/mL solution and mixed

NEGATIVE CONTROL
- Amount(s) applied: 9 g/L saline
- Concentration: 100 μL NaCl solution

POSITIVE CONTROL
- Concentration: 50 μL positive control

Duration of treatment / exposure:

4 h

Duration of post-treatment incubation (if applicable):

3 h

Number of replicates:

In this assay 2 replicates of test item, 2 replicates negative control and 2 replicates positive control were used.

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

4 h

Value:

118

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: The test item did not show significantly reduced cell viability in comparison to the negative control after 4 hours of exposure.
- Direct-MTT reduction: The test item did not interact with the MTT, therefore additional controls and data calculations were not necessary. A false estimation of viability can be precluded.
- Colour interference with MTT: No color change was observed after three hours of incubation during the check-test for possible direct MTT reduction with test item.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes; mean OD value of the two negative control tissues was 0.965.
- Acceptance criteria met for positive control: yes ; positive control result showed 2 % viability.
- Acceptance criteria met for variability between replicate measurements: yes; the difference of viability between the two tissue replicates: Negative control: 15%; Positive control: 1%; and Test item: 28%

Interpretation of results:

other: non-corrosive according to Regulation (EC) No 1272/2008

Executive summary:

In an in vitro skin corrosion human skin model test (EpiSkin) according to OECD guideline 431 and in compliance with GLP, mean cell viability was118% after 4 hours of exposure. Therefore, the test item should not be classified as corrosive.

Reference 2

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 Dec 2017 - 08 Dec 2017

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:

28 July 2015

GLP compliance:

yes (incl. QA statement)

Remarks:

Országos Gyógyszerészeti és Élelmezés-egészégügyi Intézet

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: adult donors

Source strain:

not specified

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiSkinTM Small Model (EpiSkinTMSM)
- Tissue batch number(s): 17-EKIN-049

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1°C
- Temperature of post-treatment incubation: 37 ± 1°C

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps: After the incubation time the EpiSkinTMSM units were removed and rinsed thoroughly with approximately 25 mL PBS 1x solution to remove all of the test material from the epidermal surface.

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 2 mL MTT 0.3 mg/mL solution
- Incubation time: 3 hours (± 5 min)
- Spectrophotometer: yes, plate spectrophotometer
- Wavelength: 570 nm (± 10 nm)
- Filter: no
- Linear OD range of spectrophotometer: Read out range: 0-3.5 Abs, Linearity range: 0.2136 – 3.1752

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Barrier function: 2.3 mg/mL
- Morphology: Well-differentiated epidermis consisting of a basal layer, several spinous and granular
layers and a thick stratum corneum
- Contamination: Screened for HIV-1, hepatits B and C, bacteria, yeast, and other fungi. No contaminates were detected.

NUMBER OF REPLICATE TISSUES: triplicate

PREDICTION MODEL / DECISION CRITERIA
The irritant potential was predicted from the relative mean tissue viabilities compared to the negative control. If the mean relative tissue viability is > 50% the test material is not considered an irritant. If the mean relative tissue viability is ≤ 50%, the test material is considered an irritant.

The test meets acceptance criteria if:
- mean OD570 nm of the blank is < 0.1
- mean absolute OD570 nm of the three negative control tissues is ≥ 0.6 and ≤ 1.5
- mean relative tissue viability of the three positive control tissues is ≤ 40%
- standard deviation (SD) of relative tissue viability obtained from each three concurrently tested tissues is ≤ 18%.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 10 mg

NEGATIVE CONTROL
- Amount(s) applied: 10 μL
- Concentration: 1x PBS

POSITIVE CONTROL
- Amount(s) applied: 10 μL
- Concentration: SDS 5 % aq

Duration of treatment / exposure:

15 minutes (± 0.5 min)

Duration of post-treatment incubation (if applicable):

42 hours (± 1h)

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Value:

105

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Direct-MTT reduction: The test material did not interact with the MTT.
- Colour interference with MTT: The test material did not interact with the MTT.

DEMONSTRATION OF TECHNICAL PROFICIENCY: The mean OD value of the three negative control tissues was 1.122. The mean OD value obtained for the positive control was 0.116 and this result corresponds to 10% viability when compared to the results obtained from the negative control which is in the range (0 - 40%) for the positive controls. Each calculated standard deviation value (SD) for the % viability was below 18. All validity criteria were within acceptable limits and therefore the study can be considered as valid.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes
- Acceptance criteria met for variability between replicate measurements: yes

Substance	Optical Density (OD)		Viability (%)
Test Item: Butylmonoglycol sulphate, Na-salt	1	1.150	102
	2	1.097	98
	3	1.273	114
	mean	1.174	105
	standard deviation (SD)		8.05
Negative Control: 1x PBS	1	1.097	98
	2	1.045	93
	3	1.224	109
	mean	1.122	100
	standard deviation (SD)		8.21
Positive Control: SDS (5 % aq.)	1	0.119	11
	2	0.113	10
	3	0.114	10
	mean	0.116	10
	standard deviation (SD)		0.28

Interpretation of results:

other: CLP/EU GHS criteria are not met, no classification is required according to Regulations (EC) No 1272/2008.

Conclusions:

In an in vitro skin irritation human skin model test (EpiSkin) according to OECD guideline 439 and in compliance with GLP, a cell viability of 105% was measured when compared to the untreated control. Therefore, the test item should not be classified as an irritant.

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:

5 Oct 2017 - 21 Nov 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

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:

26 July 2013

GLP compliance:

yes (incl. QA statement)

Remarks:

Országos Gyógyszerészeti és Élelmezés-egészégügyi Intézet

Species:

chicken

Strain:

other: ROSS 38

Details on test animals or tissues and environmental conditions:

SOURCE OF COLLECTED EYES
- Source: TARAVIS KFT. 9600 Sárvár, Rábasömjéni út 129. Hungary
- Number of animals: not reported
- Characteristics of donor animals: age, sex, and weight not reported
- Storage, temperature and transport conditions of ocular tissue: Head collection was performed by a slaughter house technician. Heads were removed immediately after sedation by electric current. The heads were transported at the earliest convenience for use approximately within 2 hours from collection. The ambient temperature was optimal (19.3 - 20.4ºC) during the transport.
- Time interval prior to initiating testing: All eyes used in the assay were from the same groups of eyes collected on one specific day.
- Indication of any existing defects or lesions in ocular tissue samples: The fluorescein-treated cornea was examined with hand-held slit lamp or slit lamp microscope, with the eye in the head, to ensure that the cornea was not damaged. If the cornea was in good condition, the eyeball was carefully removed from the orbit. Selected eyes were examined again with the slit lamp microscope to ensure that they were in good condition. The focus was adjusted to see clearly the isotonic saline which was 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 using the depth measuring device on the slit lamp microscope with the slit-width set at 9½, equalling 0.095 mm. 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.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.03 g

Duration of treatment / exposure:

After an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with 20 mL saline solution at ambient temperature, while taking care not to damage the cornea but attempting to remove all the residual test item if possible.

Observation period (in vivo):

The control and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Minor variations within ± 5 minutes were considered acceptable.

Number of animals or in vitro replicates:

Three test item treated eyes and three positive control eyes were used in this study. One negative control eye was treated with 30 μL saline solution.

Details on study design:

SELECTION AND PREPARATION OF ISOLATED EYES
Corneas in good condition were selected for examination. Selected eyeballs were carefully removed from the orbit by holding the nictitating membrane with surgical forceps, while cutting the eye muscles with bent scissors without cutting off the optical nerve too short. The procedure avoided pressure on the eye in order to prevent distortion of the cornea and subsequent corneal opacity. Once removed from the orbit, the eye was placed onto damp paper and the nictitating membrane was cut away with other connective tissue. The prepared eyes were kept on the wet papers in a closed box so that the appropriate humidity was maintained.

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.

NUMBER OF REPLICATES
Three test item treated eyes and three positive control eyes were used in this study. One negative control eye was treated with 30 μL saline solution.

NEGATIVE CONTROL USED : Saline solution

SOLVENT CONTROL USED: not applicable

POSITIVE CONTROL USED : Imidazole

APPLICATION DOSE AND EXPOSURE TIME : 0.03 g for 10 seconds

OBSERVATION PERIOD : 30, 75, 120, 180, and 240 minutes

REMOVAL OF TEST SUBSTANCE
- Volume and washing procedure after exposure period: After an exposure period of 10 seconds from the end of the application the cornea surface was rinsed thoroughly with 20 mL saline solution at ambient temperature .

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: Cornea opacity was measured at all time points by a hand-held slit lamp or slit lamp microscope and observations were scored on a scale from 0-4; the calculation used is Mean CO(at time t) = [FEΔCO (at time t)+ SEΔCO(at time t) + TEΔCO(at time t)]/3 where
CO = Cornea opacity;
ΔCO = Difference between cornea opacity and cornea opacity reference value;
FEΔCO = Difference between first eye cornea opacity and first eye cornea opacity reference value;
SEΔCO = Difference between second eye cornea opacity and second eye cornea opacity reference value;
TEΔCO = Difference between third eye cornea opacity and third eye cornea opacity reference value;
Mean CO = The mean corneal opacity value; at time t = Observation time at 30, 75, 120, 180 or 240 minutes after the post-treatment rinse; and at t=0 =
Reference value

- Damage to epithelium based on fluorescein retention: Fluorescein retention was measured by a hand-held slit lamp or slit lamp microscope and observations were scored on a scale from 0-3; the calculation used is Mean FR = [FEΔFR (at time t) + SEΔFR(at time t) + TEΔFR at time t)]/3 where FR = Fluorescein retention; ΔFR = Difference between fluorescein retention and fluorescein retention reference value; FEΔFR = Difference between first eye fluorescein retention and first eye fluorescein retention reference value; SEΔFR = Difference between second eye fluorescein retention and second eye fluorescein retention reference value; TEΔFR = Difference between third eye fluorescein retention and third eye fluorescein retention reference value; Mean FR = The mean fluorescein retention value; at time t = Observation time at 30 minutes after the post-treatment rinse; and at t=0 = Reference value
- Swelling: swelling was calculated through a calculation CS at time t = (CT at time t – CT at t = 0)/CT at t = 0 X 100 and Mean CS at time t = [FECS(at time t) + SECS (at time t) + TECS (at time t)]/3 where CS = Cornea swelling; CT = Cornea thickness
FECS = First eye cornea swelling; SECS = Second eye cornea swelling; TECS = Third eye cornea swelling; Mean CS = The mean percentage of corneal swelling; at time t = Observation time at 30, 75, 120, 180 or 240 minutes after the post-treatment rinse; and at t = 0 = Reference value

SCORING SYSTEM:
- Mean corneal swelling (%) : no scoring scale was used for mean corneal swelling
- Mean maximum opacity score : 0 = No opacity; 0.5 = Very faint opacity; 1 = Scattered or diffuse areas; details of the iris are clearly visible; 2 = Easily discernible translucent area; details of the iris are slightly obscured; 3 = Severe corneal opacity; no specific details of the iris are visible; size of the pupil is barely discernible; 4 = Complete corneal opacity; iris invisible
- Mean fluorescein retention score at 30 minutes post-treatment : 0 = No fluorescein retention; 0.5 = Very minor single cell staining; 1 = Single cell staining scattered throughout the treated area of the cornea; 2 = Focal or confluent dense single cell staining; 3 = Confluent large areas of the cornea retaining fluorescein

DECISION CRITERIA: used criteria established in OECD 438 guideline

Irritation parameter:

cornea opacity score

Value:

3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein retention score

Value:

2.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

percent corneal swelling

Run / experiment:

75 min

Value:

26

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

percent corneal swelling

Run / experiment:

240 min

Value:

33

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

OTHER EFFECTS:
- Visible damage on test system: yes

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes; see attached document
- Acceptance criteria met for positive control: yes; see attached document

Interpretation of results:

other: CLP/EU GHS Category 1 (H318) according to Regulation (EC) No 1272/2008

Conclusions:

In this eye irritation study (OECD 438), Butylmonoglycol sulphate, Na-salt caused ocular corrosion or severe irritation in the enucleated chicken eyes.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation

A skin corrosion study (EpiSkin™ SM) according to OECD 431 was performed with Butylmonoglycol sulphate, Na-salt (CAS 67656-24-0) (Toxi-Coop, 2017). Disks of EPISKIN were treated with test item and incubated for 4 hours. The test material was then rinsed after the exposure period and the viability of each disk was assessed by incubating the tissues for 3 hours with MTT solution. Both positive and negative controls were evaluated concurrently. The test item did not show significantly reduced cell viability in comparison to the negative control after 4 hours of exposure. The average test item treated tissue viability was 118% at 4 hours of exposure. Positive and negative controls showed the expected cell viability values within acceptable limits. All assay acceptance criteria were met, the experiment was considered to be valid. The results obtained from this in vitro skin corrosion test, using the EPISKIN model, indicated that the test item reveals no skin corrosion potential under the utilized testing conditions. In conclusion, the test item Butylmonoglycol sulphate, Na-salt can be classified as non-corrosive. 

 

An additional in vitro skin irritation study was performed with Butylmonoglycol sulphate, Na-salt (C67656 -24 -0) and was conducted in accordance to OECD 439 (Toxi-Coop, 2018). In this study, EPISKIN disks were treated and incubated with the test item for 15 minutes, rinsed, and then incubated again for an additional 42 hours. Cell viability was assessed by incubating the disks with MMT for 3 hours with MMT. SDS (5% aq.) and 1×PBS treated epidermis were used as positive and negative controls respectively. In this in vitro skin irritation test using the EPISKIN model, the test item Butylmonoglycol sulphate, Na-salt did not show significantly reduced cell viability in comparison to the negative control (mean value: 105%). Positive and negative controls showed the expected cell viability values within acceptable limits. Based on these results, the test item was considered to be non-irritant to skin.

 

In a non-GLP compliant, reliable skin irritation study (similar to OECD 404) performed with 50% solution of Butylmonoglycol sulphate, Na-salt, 3 male rabbits were exposed to 0.5 mL of the diluted test material for 4 h applied onto the shaved skin via occlusive dressing (Hüls AG, 1987). Skin reactions were evaluated according to the Draize scoring system 1, 24, 48 and 72 h post-application. No symptoms of systemic toxicity were observed in any animal during the test period and no mortality occurred during the course of the study. Mean erythema and edema scores for the mean 24/48/72 hour observation period were 0, respectively. One hour after removal of the test substance all 3 animals showed very slight erythema and 1 animal very slight edema, which all disappeared until the 24-hour reading time point. This study is considered to be a supportive study because the study was conducted with a diluted form of the substance.

 

Eye irritation

In an Isolated Chicken Eye Test (ICET) according to OECD 438 performed with Butylmonoglycol sulphate, Na-salt (CAS 67656-24-0), the test compound was applied in a single dose (30 mg/eye) onto the cornea of isolated chicken eyes (Toxi-Coop, 2017). Eyes were exposed for 10 seconds. After that the cornea surface was rinsed thoroughly and this procedure was repeated for each eye. Tested corneas were evaluated pre-treatment and at approximately 30, 75, 120, 180, and 240 minutes after the post-treatment rinse. The endpoints evaluated were corneal opacity, swelling, fluorescein retention, and morphological effects. Positive and negative controls were evaluated concurrently. Results showed that the positive control and test item were observed on the cornea surfaces at 240 min after the post-treatment rinse. Therefore, in this in vitro eye irritation study, using the Isolated Chicken Eye model with Butylmonoglycol sulphate, Na-salt, ocular corrosion and severe irritation potential was observed.

 

In a non-GLP compliant, reliable eye irritation study (similar to OECD 405) performed with 50% solution of Butylmonoglycol sulphate, Na-salt, 0.1 mL of the diluted test material was instilled into the eyes of 3 male rabbits (Hüls AG, 1987). The eyes were examined and the changes were graded according to the Draize scoring system 24, 48 and 72 h post-application. No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred during the course of the study. Twenty-four hours after test substance application 2/3 animals showed corneal opacity scored with grade 1, which was fully reversible after 48 hours. No effects on the iris were observed during the whole observation time. All 3 animals showed conjunctival redness (score 1) 1 h after test substance application. In 1/3 animals this redness was augmented at the 24-hour reading (score 2), while it remained the same in the other two animals. It was fully reversible at the 72 h reading time point in 2/3 animals and at Day 6 in 1/3 animals. Furthermore, all animals showed chemosis (score 1) after 1 h, which was fully reversible in 2/3 animals at the 48-hour reading and in 1/3 animals at the 72-hour reading. Thus, the mean values for corneal opacity, iridial and conjunctival irritation and chemosis were 0.22, 0, 0.889, and 0.556, respectively. This study is considered to be a supportive study because the study was conducted with a diluted form of the substance. Based on these results, the 50% solution of the test substance is considered to be not irritating to the eyes.

Justification for classification or non-classification

Butylmonoglycol sulphate, Na-salt does not meet the criteria for classification and labelling as skin irritant according to Regulation (EC) 1272/2008.

Butylmonoglycol sulphate, Na-salt meets the criteria for classification and labelling as Eye Damaging Category 1 with the hazard statement H318 ‘Causes serious eye damage.’ according to Regulation (EC) 1272/2008. A 50% solution of the test substance showed no eye irritation in vivo therefore a concentration limit for Eye damage Category is set to > 50%.

 There are no experimental data for respiratory tract irritation.