Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 682-872-8 | CAS number: 957787-76-7
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
While in vitro studies on skin corrosion and irritaiton did not indicate a skin irritating property of the test substance, the in vivo skin irritation test in rabbits gave a borderline positive result and the substance is classified as irritating to skin. In vitro eye irritation assays indicated that the substance should be regarded as a severe eye irritant. Further animal studies were therefore not performed.
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin irritation: in vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2011/06/07 to 2011/07/05
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 404 (Acute Dermal Irritation / Corrosion)
- Version / remarks:
- Method B4 Acute Toxicity (Skin Irritation) EC No. 440/2008
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Species:
- rabbit
- Strain:
- New Zealand White
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Leicestershire, UK
- Age at study initiation: 12 to 20 weeks old
- Weight at study initiation: 2.35 to 2.62 kg
- Housing: Individually housed in suspended cages
- Diet (e.g. ad libitum): 2930 Teklad Global Rabbit Diet ad libitum
- Water (e.g. ad libitum): Mains drinking water ad libitum
- Acclimation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17 to 23 degrees C
- Humidity (%): 30 to 70%
- Air changes (per hr): At least 15 exchanges per hour
- Photoperiod (hrs dark / hrs light): 12 hours dark/12 hours light
IN-LIFE DATES: From: 2011/06/07 To: 2011/07/05 - Type of coverage:
- semiocclusive
- Preparation of test site:
- other: Clipped with veterinary clippers
- Vehicle:
- unchanged (no vehicle)
- Controls:
- no
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 ml
- Concentration (if solution): undiluted - Duration of treatment / exposure:
- One animal initially treated: three suitable sites with test article applied. One patch removed at either 3 minutes, 1 hour or 4 hours after application. Any residual test article was removed by gentle swabbing with cotton wool soaked in distilled water.
After consideration of the reactions in the first animal, an additional two animals were treated with 0.5 ml of test item. One patch was applied to the back of each of the two animals and allowed to remain in contact for 4 hours. - Observation period:
- Observations and scoring of the skin reactions were made immediately after removing the patch, at 1, 24, 48 and 72 hours. Additional observations were made on days 7 and 14 to assess the irreversibility of the lesions.
Body weights were recorded on the day of dosing and at the end of the observation period - Number of animals:
- Three male New Zealand White rabbits
- Details on study design:
- TEST SITE
- Area of exposure: under 2.5 cm square cotton gauze patch.
- Type of wrap if used: Strip of surgical adhesive tape, with trunk wrapped in an elasticized corset.
REMOVAL OF TEST SUBSTANCE
- Washing (if done): Gentle washing with cotton wool soaked in distilled water.
- Time after start of exposure: Immediately after patch removal (3 minutes, 1 hour or 4 hours).
SCORING SYSTEM: Draize J H (1959)
Erythema and Eschar Formation Value
No erythema 0
Very slight erythema 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema to eschar formation
preventing grading of erythema 4
Oedema formation
No oedema 0
Very slight oedema 1
Well-defined oedema 2
Moderate to severe oedema 3
Severe oedema 4 - Irritation parameter:
- primary dermal irritation index (PDII)
- Basis:
- mean
- Time point:
- other: 24 and 72 hours
- Score:
- ca. 4
- Reversibility:
- fully reversible within: 14 days
- Remarks on result:
- other: 4 hour exposure, all three animals
- Irritation parameter:
- erythema score
- Basis:
- animal #1
- Remarks:
- mean 24, 48, 72 h
- Time point:
- other: 24, 48, 72 h
- Score:
- 2
- Max. score:
- 2
- Reversibility:
- fully reversible within: 14 days
- Remarks on result:
- other: 4 h exposure, light brown discoloration of the epidermis at 48 and 72 h, loss of skin elasticity and flexibility at 72 h, hardened dark coloured and undulating scab at 72 h and 7d.
- Irritation parameter:
- erythema score
- Basis:
- animal #2
- Remarks:
- mean 24, 48, 72 h
- Time point:
- other: 24, 48, 72 h
- Score:
- 2
- Max. score:
- 2
- Reversibility:
- fully reversible within: 14 days
- Remarks on result:
- other: 4 h exposure, light brown discoloration of the epidermis at 48 and 72 h, moderate desquamation at day 7, no reading at day 14.
- Irritation parameter:
- erythema score
- Basis:
- animal #3
- Remarks:
- mean 24, 48, 72 h
- Time point:
- other: 24, 48, 72 h
- Score:
- 2
- Max. score:
- 2
- Reversibility:
- fully reversible within: 7 days
- Remarks on result:
- other: 4 h exposure, light brown discoloration of the epidermis at 24, 48 and 72 h, hardened dark coloured scab at 24, 48 and 72 h, moderate desquamation on day 7, no reading on day 14.
- Irritation parameter:
- edema score
- Basis:
- animal #1
- Remarks:
- mean 24, 48, 72 h
- Time point:
- other: 24, 48, 72 h
- Score:
- 2
- Max. score:
- 2
- Reversibility:
- fully reversible within: 14 days
- Remarks on result:
- other: 4 h exposure
- Irritation parameter:
- edema score
- Basis:
- animal #2
- Remarks:
- mean 24, 48, 72 h
- Time point:
- other: 24, 48, 72 h
- Score:
- 1.67
- Max. score:
- 2
- Reversibility:
- fully reversible within: 7 days
- Remarks on result:
- other: 4 h exposure
- Irritation parameter:
- edema score
- Basis:
- animal #3
- Remarks:
- mean 24, 48, 72 h
- Time point:
- other: 24, 48, 72 h
- Score:
- 2.33
- Max. score:
- 3
- Reversibility:
- fully reversible within: 7 days
- Remarks on result:
- other: 4 h exposure
- Irritant / corrosive response data:
- See Any other information on results section for data used in calculating primary irritation index.
- Other effects:
- All three animals showed expected body weight changes over the course of the study. Day 0 to Day 14 weight change in animal 1 was 0.23 kg. Animals 2 and 3 gained 0.08 and 0.17 kg over Days 0 to Day 7 of the study.
- Interpretation of results:
- Category 2 (irritant)
- Remarks:
- Migrated information Criteria used for interpretation of results: EU
- Conclusions:
- The test item produced a primary irritation index of 4.0 and was classified as a moderate irritant to rabbit skin according to the Draize classification, when applied under a semi-occlusive dressing for 4 hours. 3 minute and 1 hour exposure of the test item to the intact skin of one rabbit produced no corrosive effects.
According to CLP criteria the the scores for erythema and/or edema are below the score of 2.3 in at least 2 of 3 animals after 4 h of exposure. The effects were reversible within the 14 day observation period. However as skin discoloration and scab formation occurred during the observation period, a classification as category 2 irritant is proposed.
According to Dir. 67/548/EC and adaptations the scores for erythema and/or edema were 2 or greater in 2 of three animals and the substance is consequently classified as irritant to skin R38. - Executive summary:
A rabbit dermal irritation study conducted by the method of OECD 404 was conducted using New Zealand White Rabbits. A single animal showed no corrosive effects from a 3 -minute or a 1 hour semi-occluded application of test article to intact skin. Scores for erythema increased from 24 to 72 hours, with maximum scores of 2 at the 72 hour reading. Blanching of the skin was noted at 72 hours in the 3 minute and 72 hour sites. Crusting was seen at 7 days after the 1 hour exposure, but other than glossy skin at the 3 minute exposure site, the skin had returned to normal by 14 days. Oedema readings increased to a maximum of 2 at 72 hours in the 3 minute and 1 hour exposure sites. Edema readings could not be assessed at 7 days in the 1 hour exposure site. Readings for edema were normal at 14 days in both exposure sites.
In the 4 hour exposure portion of the experiment, an additional 2 rabbits were added to the original for a total of 3 rabbits. A single 4 hour exposure of the test item to intact skin on these 3 animals produced well-defined erythema and slight to moderate edema. Maximum scores for erythema of 2 were seen for all rabbits at the 24 to 72 hour readings. Other skin readings observed were brown discoloration of the epidermis, hardened dark brown colored scab, loss of skin elasticity and flexibility, undulating scab, modreate desquamation and crust formation than hindered accurate evaluation of edema and erythema in one rabbit at 7 days. Two rabbits had normal readings except for some desquamation at 7 days, and the thrid rabbit had normal readings at 14 days. In t he opinion of the Study Director, these observations did not indicate corrosion.
Based on a Primary Irritation Index of 4.0, the test article was classified as a moderate irritant to rabbit skin according to the Draize classification scheme.
According to CLP (Reg. 1907/2006 and amendments) criteria the the scores for erythema and/or edema are below the score of 2.3 in at least 2 of 3 animals after 4 h of exposure. The effects were reversible within the 14 day observation period. However as skin discoloration and scab formation occurred during the observation period, as classification as category 2 irritant is proposed.
According to Dir. 67/548/EC and adaptations the scores for erythema and/or edema were 2 or greater in 2 of three animals and the substance is consequently classified as irritant to skin R38.
Reference
|
|
Skin reactions after 3 min and 1 hour of exposure (one male animal):
|
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation
- Remarks:
- in vitro
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 26 August 2011
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Followed established Guideline and GLP requirements
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 405 (Acute Eye Irritation / Corrosion)
- Version / remarks:
- In vitro test in step wise procedure of Guideline 405
- Qualifier:
- according to guideline
- Guideline:
- other: OECD No. 437 (2009) "Bovine Corneal Opacity and Permeability Assay"
- GLP compliance:
- yes (incl. QA statement)
- Species:
- other: bovine corneas
- Details on test animals or tissues and environmental conditions:
- TEST ANIMALS
- Source: local abbatoir
- Age at study initiation: adult cattle, eyes obtained from freshly slaughtered animals
ENVIRONMENTAL CONDITIONS
- Temperature (°C): eyes transported in Hanks' Balanced Salt Solution supplemented by Penicillin/Streptomycin and placed on ice packs. Eyes refrigerated on arrival to lab, and used within 24 hours. - Vehicle:
- unchanged (no vehicle)
- Controls:
- yes
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.75 mL test item or control item applied to each cornea.
- Concentration (if solution): Negative control was 0.9% sodium chloride solution. - Duration of treatment / exposure:
- After test item or control item applied to cornea in the BCOP holder and the holder tilted to assure uniform applicaiton to the cornea, the holder containing the cornea was incubated at 32 +/- 1 degrees C for 10 minutes. The cornea was then rinsed three times with complete Minimum esssential medium (MEM) containing phenol red before a final rinse with complete MEM.
- Observation period (in vivo):
- A pre-treatment opacity reading of each cornea was taken before the exposure period. A post-treatment opacity reading was taken and each cornea observed visually after the rinsing with complete MEM. The holders were then incubated at 32 +/- 1 degrees C for 120 minutes +/- 10 minutes. After that incubation, a final opacity reading was taken and each cornea visually observed.Following the final opacity reading, the medium was removed from the anterior chamber and replaced with 1 ml sodium fluorescein (4 mg/ml). The holders were incubated for 90 minutes at 32 +/- 1 degrees C. Medium from the posterior chamber was used for optical density reading (a sample of 360 ul represented each cornea.
- Number of animals or in vitro replicates:
- Three corneas each for test item, negative control and positive control
- Details on study design:
- REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, after 10 minutes, the corneas were rinsed with MEM plus phenol red three times before a final rinse with complete MEM
- Time after start of exposure: 10 minutes of exposure
SCORING SYSTEM: Opacity reading using calibrated opacitometer; permeability measured by optical density at 492 nm;
TOOL USED TO ASSESS SCORE: Calibrated opacitometer; 96 well plate for measuring optical density at 429 nm. - Irritation parameter:
- other: In Vitro Irritation Score (IVIS)
- Basis:
- other: mean value for treated cornea group
- Score:
- ca. 66
- Remarks on result:
- other: The IVIS score was greater then 55, thus the test item is a severe irritant or corrosive
- Irritant / corrosive response data:
- See tables in results section for individual corneal and mean treatment values for opacity, permeability, corrected values of each, and the resulting IVIS scores.
- Interpretation of results:
- highly irritating
- Remarks:
- Migrated information Criteria used for interpretation of results: other: Criteria in OECD No. 437
- Conclusions:
- Following assessment of all endpoint data in the Bovine Corneal Opacity and Permeability Assay, the test item was considered to have the potential to cause severe ocular irritancy in vivo. The in vivo eye irritation study is therefore not needed.
According to CLP (Reg. 1907/2006 and amendments) the substance is classified as causing irreversible eye effects (category 1) based on this in vitro result.
(Reg. 1907/2006 and amendments) the substance is classified as posing the risk of serious damage to eyes (R41). - Executive summary:
Following assessment of all endpoint data in the Bovine Corneal Opacity and Permeability Assay, the test item was considered to have the potential to cause severe ocular irritancy in vivo. The in vivo eye irritation study is therefore not needed. The In Vitro Irritancy Score, calculated on changes in opacity and permeability (optical density), was 66.0 for the test item compared to 67.4 for the positive control item (ethanol). According to the OECD Guideline No. 437, an IVIS of 55 or greater is interpreted as the test item has the potential for severe ocular irritancy in vivo.
According to CLP (Reg. 1907/2006 and amendments) the substance is classified as causing irreversible eye effects (category 1) based on this in vitro result.
(Reg. 1907/2006 and amendments) the substance is classified as posing the risk of serious damage to eyes (R41).
- Endpoint:
- eye irritation
- Remarks:
- other: in vitro human cornea model
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- 2011-04-12 to 2011-04-14
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: Follwos accepted scientific principles and conducted under GLP
- Qualifier:
- according to guideline
- Guideline:
- other: Van Goethem F, et al,"Prevalidation of a new in vitro reconstituted human cornea model to assess the eye irritating potential of chemicals," Toxicology in vitro, 20 (2006)
- Principles of method if other than guideline:
- The SkinEthic HCE model consists of transformed human corenal epithelial cells of the cel line HCE that form a corneal epithelial tissue (mucosa) devoid of stratum corneum, resembling, histologically, the mucosa of the human eye. The test article is applied to the culture surface at the air interface so that undiluted or end use dilutions can be teste directly. the model consists of an airlifted, living corneal tissue construct, produced in polycarbonate inserts in serm-free and chemically defined medium. The test is based on the hypothesis that irritant chemicals are able to penetrate the stratum corneum of the HCE model and are sufficiently cytotoxic to cause cell death in underlying cell layers, Cytotoxicity was determined by reduction of MTT to formazan by living cells in the test item treated tissues (quantitative measurement of cell viability) relative to the negative control). Measurement of the optical density (OD540) quantified the color change of the MTT to formazan. One tissue per treatment group was retained for possible histopatholoty.
- GLP compliance:
- yes (incl. QA statement)
- Species:
- other: transformed human corneal epithelial cells
- Strain:
- other: Cell line HCE (LSU Eye Center, New Orleans, USA
- Details on test animals or tissues and environmental conditions:
- TEST System: SKINETHIC HCE Model
- Source: SkinEthic Laboratories, Nice, France
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 37 degrees C, 5% CO2 in air storage conditions for receipt and overnight incubation of Day 6 cultures
Study DATES: From: 12 April 2011 To: 14 April 2011 - Vehicle:
- unchanged (no vehicle)
- Controls:
- other: negative and positive control groups
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): MTT direct reduction assessment: 30 ul test item to 1 ml of 0.5 mg/mL MTT solution
Main assay: 30 ul test item or control item applied to triplicate tissues for 10 minutes
- Concentration (if solution): test item as supplied
SDS positive control prepared as 2% w/v in sterile water
Solution A negative control used as supplied
Control substances:
Negative Solution A as supplied by Test kit
Positive Sodium dodecyl sulphate (SDS) 2%w/v prepared in distilled water - Duration of treatment / exposure:
- Ten minutes exposure of tissues
- Observation period (in vivo):
- Main test: following exposure, and tissue rinsing, each tissue was exposed to the MTT solution and incubated for 3 hours. At the end of the incubatin period, the tissues were examined, and the degree of MTT staining assessed (qualitative evaluation). Tissues were then mived to the MTT extraction plates, isopropanol applied, and the plates covered with foil and allowed to stand at room temperature overnight to extract the formazan crystals. At the end of the extractin period, extraction fluids were withdrawn and optical density (quantitative assessment of viability) made.
- Number of animals or in vitro replicates:
- Three tissues for treatment group - 1 tissue retained for possible histopathology
- Details on study design:
- REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes, with Dulbecco's Phosphate Buffered Saline with Ca++ and Mg++,
- Time after start of exposure: to minutes
SCORING SYSTEM: OD540 readings used to calculate relative mean tissue viability of treated group
Relative mean tissue viability (%) = (mean OD540 of test item/mean OD540 of negative control) x 100
TOOL USED TO ASSESS SCORE: Anthos 2001 microplate reader
Classification:
Relative mean tissue viability Prediction
Tissue viability < 60% Irritant (I)
Tisue viability =/< 60% Non-irritant
Criteria for a Valid Test:
Positive control relative mean tissue viability < 60% relative to negative control treated tissues
MTT direct test shows chemical does not directly reduce MTT - Irritation parameter:
- other: relative mean tissue viability (%)
- Basis:
- other: mean viability of treated compared to control tissues
- Time point:
- other: 10 minute exposure plus time for MTT expsoure, formazan extraction and OD540 readings
- Score:
- ca. 52.6
- Irritant / corrosive response data:
- See Any Other Information Section
- Interpretation of results:
- irritating
- Remarks:
- Migrated information Criteria used for interpretation of results: other:
- Conclusions:
- The test article was considered to be irritating to the eye based on the relative cell viability of < 60% compared to control tissues in the SKINETHIC Human corneal Epithelial Model assay with 10 minute exposure.
- Executive summary:
The purpose of the test was to evaluate eye irritation potential of the test article using the SKINETHIC reconstructed human corneal model (HCE, SkinEthic Laboratories, Nice, France) after a treatment period of 10 minutes. The principle of the assay was based on measurement of cytotoxicity in reconstructed human corneal cell cultures following topical exposure to test item by means of colorimetric MTT reduction assay. Cell viability is measured by enzymatic reduction of the yellow MTT tetrazolium salt (3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to a blue formazan salt (within the mitochondria of viable cells) in the test item treated tissues relative to negative controls.
The relative mean viability of the test article treated tissues was 52.6% after the 10 minute exposure. Quality criteria for the acceptance of the results were satisfied. The test article was considered to be irritant.
Referenceopen allclose all
Individual and Mean Corneal Opacity Measurements
Opacity |
||||
Treatment |
Cornea Number |
Pretreatment/ Postreatment/Post Incubation |
Post incubation - Pretreatment |
Corrected Value |
Negative Control |
1 |
1 1 2 |
1 |
|
2 |
1 1 1 |
0 |
||
3 |
2 2 3 |
1 |
||
Mean |
0.7 |
|||
Positive Control |
4 |
2 28 28 |
26 |
25.3 |
5 |
1 27 27 |
26 |
25.3 |
|
6 |
2 25 25 |
23 |
22.3 |
|
Mean |
24.3 |
|||
Test Item |
7 |
3 5 9 |
6 |
5.3 |
8 |
1 10 13 |
12 |
11.3 |
|
9 |
1 4 18 |
17 |
16.3 |
|
Mean |
11.0 |
Individual and Mean Permeability Measurements and Resulting IVIS
Permeability (Optical Density, OD) |
|||||
Treatment |
Cornea Number |
Corrected Value |
In vitro Irritancy Score (IVIS) |
||
Negative Control |
1 |
0.057 |
|||
2 |
0.127 |
||||
3 |
0.056 |
||||
Mean |
0.080 |
1.9 |
|||
Positive Control |
4 |
2.860 |
2.780 |
||
5 |
2.650 |
2.570 |
|||
6 |
3.350 |
3.270 |
|||
Mean |
2.873 |
67.4 |
|||
Test Item |
7 |
4.545 |
4.465 |
||
8 |
3.665 |
3.585 |
|||
9 |
3.025 |
2.945 |
|||
Mean |
3.665 |
66.0 |
Corneal Epithelium Post Treatment and Post Incubation
Observation |
||||
Treatment |
Cornea Number |
Post Treatment |
Post Incubation |
|
Negative Control |
1 |
Clear |
Clear |
|
2 |
Clear |
Clear |
|
|
3 |
Clear |
Clear |
|
|
Positive Control |
4 |
Cloudy |
Cloudy |
|
5 |
Cloudy |
Cloudy |
|
|
6 |
Cloudy |
Cloudy |
|
|
Test Item |
7 |
Clear |
Cloudy |
|
8 |
Cloudy |
Cloudy |
|
|
9 |
Clear |
Cloudy |
|
Mean OD540 Values and Percentage Viabilities for the Negative Control, Positive Control and Test Items
Item |
OD540of tissues |
Mean OD540(triplicate tissues) |
Relative mean viability (%) |
Negative Control Item |
0.895 0.879 |
0.887 |
100* |
Positive Control Item |
0.222 0.171 |
0.197 |
22.2 |
Test Item |
0.517 0.417 |
0.467 |
52.6 |
* mean viability of the negative control tissues is set at 100%
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Additional information
The skin and eye irritation properties of the test material were investigated using a tiered testing strategy. From physical chemical and structural considerations an irritating or corrosive effect could not be excluded, but also not positively concluded. Therefore in vitro studies for skin and eye irritation/corrosion were performed.
The test item was assessed for skin corrosivity potential using the Corrositex assay. The method designed was compatible with OECD Guidelines for the Testing of Chemicals, No. 435 “In vitro Membrane Barrier Method for Skin Corrosion.” The assay consisted of a qualification screen to determine if the chemical could be detected by the Chemical Detection System, a categorization screen to determine test items with high or low acid/alkaline reserve and the definitive test. The test article did qualify for the detection system and was determined to be a Category 1 chemical (high acid/alkaline reserve). No breakthrough of the membrane was noted after 4 hours. It was concluded that the test article was non-corrosive under the conditions of the test.
Therefore the skin irritation potential of the test article was examined in a second in vitro test using the EPISKIN reconstructed human epidermis model after a treatment period of 15 minutes and a post-exposure incubation period of 42 hours. The principle of the assay was based on measurement of cytotoxicity in reconstructed human epidermal cultures following topical exposure to test item by means of the colorimetric MTT reduction assay. The relative mean viability of the test article treated tissues was 80.7% after the 15 minute exposure period. Quality criteria for the acceptance of the results were satisfied. The test article was considered to be non-irritant.
Following the negative in vitro result, a skin irritation study in rabbits was performed with the substance according to OECD 404 using New Zealand White Rabbits. A single animal showed no corrosive effects from a 3 -minute or a 1 hour semi-occluded application of test article to intact skin. Scores for erythema increased from 24 to 72 hours, with maximum scores of 2 at the 72 hour reading. Blanching of the skin was noted at 72 hours in the 3 minute and 72 hour sites. Crusting was seen at 7 days after the 1 hour exposure, but other than glossy skin at the 3 minute exposure site, the skin had returned to normal by 14 days. Oedema readings increased to a maximum of 2 at 72 hours in the 3 minute and 1 hour exposure sites. Edema readings could not be assessed at 7 days in the 1 hour exposure site. Readings for edema were normal at 14 days in both exposure sites. In the 4 hour exposure portion of the experiment, an additional 2 rabbits were added to the original for a total of 3 rabbits. A single 4 hour exposure of the test item to intact skin on these 3 animals produced well-defined erythema and slight to moderate edema. Maximum scores for erythema of 2 were seen for all rabbits at the 24 to 72 hour readings. Other skin readings observed were brown discoloration of the epidermis, hardened dark brown colored scab, loss of skin elasticity and flexibility, undulating scab, modreate desquamation and crust formation that hindered accurate evaluation of edema and erythema in one rabbit at 7 days. Two rabbits had normal readings except for some desquamation at 7 days, and the thrid rabbit had normal readings at 14 days. In the opinion of the Study Director, these observations did not indicate corrosion. Although the results indicate a borderline case with regard to classification as irritant it is concluded that the substance should be considered as irritating to skin and classified accordingly.
The eye irritation potential was concluded based on the results of 2 in vitro experiments.
The first experiment was based on the SKINETHIC reconstructed human corneal model (HCE, SkinEthic Laboratories, Nice, France) after a treatment period of 10 minutes. The principle of the assay was based on measurement of cytotoxicity in reconstructed human corneal cell cultures following topical exposure to test item by means of colorimetric MTT reduction assay.
The relative mean viability of the test article treated tissues was 52.6% after the 10 minute exposure. Based on this result the test article was considered to be irritant to eyes.
The second experiment was the Bovine Corneal Opacity and Permeability Assay. The In Vitro Irritancy Score, calculated on changes in opacity and permeability (optical density), was 66.0 for the test item compared to 67.4 for the positive control item (ethanol). According to the OECD Guideline No. 437, an IVIS of 55 or greater is interpreted as a potential for severe ocular irritancy in vivo. Based on this study it is concluded that the substance may cause irreversible eye effects.
Effects on skin irritation/corrosion: irritating
Effects on eye irritation: highly irritating
Justification for classification or non-classification
Skin irritation: The substance is classified based on the results of the in vivo skin irritation study in rabbits.
According to CLP (Reg. 1907/2006 and amendments) criteria the the scores for erythema and/or edema are below the score of 2.3 in at least 2 of 3 animals after 4 h of exposure. The effects were reversible within the 14 day observation period. However as skin discoloration and scab formation occurred during the observation period, a classification as category 2 irritant is proposed.
According to Dir. 67/548/EC and adaptations the scores for erythema and/or edema were 2 or greater in 2 of three animals and the substance is consequently classified as irritant to skin R38.
Eye irritation: Based on the result of an in vitro Bovine Corneal Opacity and Permeability Assay, the substance is classified as causing irreversible eye effects (category 1) according to CLP (Reg. 1907/2006 and amendments).
According to Dir. 67/548/EC and adaptations
the substance is classified as posing the risk of serious damage to eyes (R41).
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.