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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 267-499-7 | CAS number: 67874-71-9
- 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
Skin corrosion/irritation: irritating (OECD 435 and OECD 439; GLP)
Eye irritation: causes serious eye damage (OECD 405; GLP)
Key value for chemical safety assessment
Skin irritation / corrosion
Link to relevant study records
- Endpoint:
- skin corrosion: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2018-05-09
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)
- Version / remarks:
- 2015-07-28
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- signed 2018-04-26
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: -15°C to 40°C - Test system:
- artificial membrane barrier model
- Source species:
- other: not specified
- Cell type:
- other: synthetic macromolecular bio-barrier
- Cell source:
- other: not specified
- Source strain:
- not specified
- Details on animal used as source of test system:
- not applicable
- Justification for test system used:
- The CORROSITEX™ Assay is a standardized, quantitative in vitro test for skin corrosivity and has been validated by the ECVAM for testing acids, bases and their derivatives (ECVAM, 2000)*. The bio-barrier membrane is constructed to have physico-chemical properties similar to rat skin.
Reference:
- ECVAM (2000) ESAC statement on the application of the Corrositex® assay for skin corrosivity testing - Vehicle:
- unchanged (no vehicle)
- Details on test system:
- SOURCE AND COMPOSITION OF MEMBRANE BARRIER USED
- Was the Corrositex® test kit used: yes (lot no. CT120516; supplier: Invitro International; Irvine, CA 92614)
- Components: a synthetic macromolecular bio-barrier and a chemical detection system (CDS)
- Apparatus and preparation procedures: the preparation was completed at least 2 hours prior to running tests. The entire content of the BIOBARRIER diluent was added to the vial of BIOBARRIER matrix powder. The vial was heated to 68°C (± 1°C) in a water bath under smooth agitation. After complete dissolution (approx. 20 min.) the solution was allowed to sit for 5 min. to allow any air bubbles to rise to the surface. 200 µL of the BIOBARRIER were pipetted into each membrane disc. The BIOBARRIERS were set on the tray and kept in the cold (2 - 8°C) for at least two hours.
WAS THE COMPATIBILITY TEST PERFORMED: yes
In order to test whether the test system is suitable for the test item, 150 µl of the test substance was added to the Qualify test tube. The vial was shaken to allow dissolution of the test substance and let stand for one minute. If the colour or consistency of the CDS changes at the sample/testing fluid interface, the test material is qualified for the assay. If no reaction is observed within five minutes, the sample is not qualified for the CORROSITEX TM assay.
WAS THE TIMESCALE CATEGORY TEST PERFORMED: yes
This step established the category of cut-off times for the sample. 150 µl of the test substance was added to the tubes labelled Tube A and Tube B. The vial was shaken to allow dissolution of the test substance. In case a colour change was observed in either of the tubes and colour was matched to the corresponding colour charts on the CORROSITEX™ Testing Protocol Poster. Test materials having high acid/alkaline reserves are defined as Category 1 materials, while those with low acid/alkaline reserves are defined as Category 2 materials. If no colour change had been observed in either tube, CONFIRM reagent was added to Tube B. After shaking, the resulting colour was matched to the colour chart on the CORROSITEX™ Testing Protocol Poster. If the test item has a strong inherent colour or shows other characteristics impairing a clear categorization according to the colour chart, the pH value can be measured in tubes A and B and is used to confirm/determine the category of the test item, according to the Corrositex® Reference Manual (1995)*.
TEMPERATURE USED DURING TREATMENT: room temperature (17 - 25 °C)
METHOD OF DETECTION
- Chemical or electrochemical detection system: chemical detection system (CDS)
METHOD OF APPLICATION (CLASSIFICATION TEST):
The CDS vials were warmed to room temperature (17 - 25˚C) before using. Four vials were utilized for test item sample replicate testing. One vial was utilized for a positive control sample and another vial for a negative control. Lastly, one vial served as a CDS colour control. One BIOBARRIER disc was added on top of the first vial (discs were not longer in the vial than two minutes before adding the test samples).
500 µl of the test item was applied evenly on the top of the BIOBARRIER disc and starting time was recorded. This step was repeated for the remaining vials, staggering each start time by e.g. 10 seconds (but not longer than 2 minutes). The start time difference for each vial was subtracted from the final time to determine the net response time. As soon as a reaction had been observed, the time was recorded.
NTERPRETATION OF THE RESULTS:
For Category 1 substances, test chemicals were categorized as non-corrosive in case no colour change occurs after 240 minutes. For Category 2 substances, test chemicals were categorized as non-corrosive in case no colour change occurs after 60 minutes. The start time difference for each vial was subtracted from the final time to determine the net response time.
The time (in minutes) elapsed between application and barrier penetration for the test substance was recorded in tabular form as individual replicate data.
The mean time (± standard deviation) of the four sample replicates to activate the CDS was calculated and reported in tabular form. Using the table as shown in the field "Any other information on materials and methods incl. tables" below, the test item was categorised.
TEST ACCEPTANCE CRITERIA:
The test meets acceptance criteria if:
- test item qualifies in qualification test
- positive control activates CDS > 3 - 60 min.
- negative control activates CDS not before 60 min.
The exact breakthrough time of the positive control should be determined to demonstrate, that the response is in the acceptable historical range of breakthrough times for the positive control (mean ± 2 - 3 standard deviations).
*Reference:
- InVitro International (1995), Corrositex® Reference Manual - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 500 µL of the test item
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 500 µL of citric acid (10%) in aqua dest.
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 500 µL of phosphoric acid (85 %) - Duration of treatment / exposure:
- 60 minutes
- Duration of post-treatment incubation (if applicable):
- not applicable
- Number of replicates:
- Test item: quadruplicates
Negative control: single measurement
Positive control: single measurement - Irritation / corrosion parameter:
- penetration time (in minutes)
- Value:
- 0
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: Colour change or structural change was not observed up to 60 minutes (treatment period)
- Other effects / acceptance of results:
- QUALIFICATION TEST
The test substance was compatible with the CORROSITEX™ Assay, as assessed in the qualification step. The categorization step and the classification step could be performed.
CATEGORIZATION TEST
A direct colour change was not observed. CONFIRM reagent was added to tube B and the category was read from the CORROSITEX™ colour chart. The chemical has been categorized to timescale category 2.
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: negative control did not activate the CDS before 60 minutes (> 60 minutes).
- Acceptance criteria met for positive control: positive control activated the CDS between 3 - 60 minutes (26.18 min).
Please also refer to the field "Any other information on results incl. tables" below. - Interpretation of results:
- GHS criteria not met
- Conclusions:
- According to the Regulation (EC) No 1272/2008 and subsequent adaptations, the test item bismuth tris(2-ethylhexanoate) is not corrosive to the skin.
- Endpoint:
- skin irritation: in vitro / ex vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2018-04-25 to 2018-04-27
- 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:
- 2015-07-28
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- other: MatTek Corporation Protocol for: In Vitro EpiDermTM Skin Irritation Test (EPI-200-SIT) For use with MatTek Corporation’s Reconstructed Human Epidermal Model EpiDerm (EPI-200-SIT)
- Version / remarks:
- 2014-11-07
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- signed 2015-06-05
- Specific details on test material used for the study:
- STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: -15°C to 40°C - Test system:
- human skin model
- Source species:
- human
- Cell type:
- other: normal, human epidermal keratinocytes
- Cell source:
- other: humans
- Source strain:
- other: not applicable
- Details on animal used as source of test system:
- not applicable
- Justification for test system used:
- This test uses the EpiDerm™ reconstructed human epidermis model (MatTek) which consists of normal human-derived epidermal keratinocytes (NHEK) and therefore represents in vitro the target organ of the species of interest and closely resembles the biochemical and physiological properties of the upper parts of the human skin, i.e. the epidermis.
- Vehicle:
- unchanged (no vehicle)
- Details on test system:
- RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDermTM (EPI-200-SIT; MatTek)
- Tissue lot number: 25899
TEST FOR MTT INTERFERENCE
- to check the non-specific MTT-reducing capability of the test item 25 mg of the test item were mixed per 1 mL MTT medium and incubated for 60 min at 37 ± 1 °C in the incubator (5 % CO2, 95 % RH).
- untreated MTT medium was used as control.
- after incubation verification of the colour by the unaided eye.
TEST FOR COLOUR INTERFERENCE
- to check the colouring potential of the test item 25 mg of the test item were mixed per 300 µL aqua dest. and per 300 µL isopropanol each in a transparent recipient and incubated at 37 ± 1°C for 60 min (5 % CO2, 95 % RH).
- after incubation verification of the colour by the unaided eye
TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 ± 1 °C for 35 ± 1 minutes followed by incubation at room temperature until the 60 ± 1 minute treatment period was completed
- Temperature of post-treatment incubation: 37 ± 1 °C
REMOVAL OF TEST MATERIAL AND CONTROLS
- after the end of the treatment period the tissues were washed 15 times with DPBS.
- subsequently, the inserts were submerged three times in DPBS and shaken to remove rests of the test item.
- then inserts were rinsed once from the inside and the outside with sterile DPBS.
- inserts were placed in prepared 6-well plates containing pre-warmed fresh assay medium per well.
- plates were post-incubated at 37 ± 1 °C, 5.0% CO2, humidified to 95%, for 24 ± 2 h. Following this incubation the tissues were transferred to new wells containing fresh assay medium and incubated for additional 18 ± 2 h.
MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL (300 µL/well)
- Incubation time: 3 hours ± 5 minutes
- Extraction of formazan: after the MTT incubation period, the tissues were rinsed three times with DPBS and allowed to dry. The tissues were transferred into 12-well plates and immersed in 2 mL isopropanol, sealed to inhibit evaporation. Extraction was carried out protected from light at room temperature at least for 2 hours with shaking on a plate shaker.
Before using the extracts, the plate had been shaken for at least 15 minutes on a plate shaker and the inserts were pierced with an injection needle. The extract was pipetted up and down 3 times before 2 x 200 µL aliquots per each tissue were transferred into a 96-well plate. Optical density (OD) was measured with a filter band without reference wavelength in a plate spectrophotometer using isopropanol as a blank.
- Wavelength: 570 nm
- Filter bandwidth: maximum ± 30 nm
FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: tissues pass analysis for tissue viability
- Barrier function: tissues pass analysis for tissue functionality
- Morphology: presence of a functional stratum corneum, a viable basal cell layer, and intermediate spinous and granular layers.
- Contamination: absence of bacteria, yeast, and other fungi (long term antibiotic, antimycotic free culture) as well as absence of HIV1-virus, Hepatitis B virus and Hepatitis C virus
Please also refer to the field "Attached background material" below.
PREDICTION MODEL / DECISION CRITERIA
The mean optical density (OD) of the three negative control tissues was calculated after blank correction. This value corresponded to 100 % tissue viability in the current test. For each individual tissue treated with the test item or the positive control, the individual relative tissue viability is calculated according to the following formula:
Relative viability (%) = [(mean ODtest item / positive control) / ODmean of negative control] * 100
For the test item and the positive control the mean relative viability ± relative standard deviation of the three individual tissues were calculated and used for classification according to the following prediction model:
Irritant potential of the test item was predicted from the relative mean tissue viabilities compared to the negative control tissues concurrently treated with DPBS.
The test item is identified as requiring classification and labelling according to UN GHS/ EU CLP “Category 2” or “Category 1", if the tissue viability after exposure and post-incubation is less or equal to 50%. Further testing is required to resolve between UN GHS categories 1 and 2 and decide on the final classification of the test substance. The test substance may be considered as non-irritant to skin in accordance with regulation EC 1272/2008 and UN GHS “No Category”, if the tissue viability after exposure and post-treatment incubation is more than 50%. - Control samples:
- yes, concurrent negative control
- yes, concurrent positive control
- Amount/concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 µL (47 µL/cm²) of the test item
Prior to treatment, all EpiDerm™ tissues were gently blotted to remove moisture. The test item was applied undiluted. 30 µL (47 µL/cm²) of the test item were dispensed directly atop the EpiDerm™ tissue. The test item was gently spread to match size of the tissue using a bulb-headed Pasteur pipette. After the washing steps, a small amount of the test material could not be removed from the edge of the tissue surface
NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL DPBS
POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL of 5 % SDS solution - Duration of treatment / exposure:
- 60 ± 1 minute
- Duration of post-treatment incubation (if applicable):
- approx. 42 hours
- Number of replicates:
- triplicates
- Irritation / corrosion parameter:
- % tissue viability
- Remarks:
- (mean)
- Value:
- 4.8
- Vehicle controls validity:
- not examined
- Negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other:
- Remarks:
- After the washing steps, a small amount of the test material could not be removed from the edge of the tissue surface.
- Other effects / acceptance of results:
- TEST FOR MTT INTERFERENCE
The mixture of 30 µL test item per 1 mL MTT medium showed no reduction of MTT compared to the solvent. The mixture did not turn blue/purple. Therefore, the test item did not directly reduce MTT (NSMTT equalled 0%) and a functional test with freeze-killed tissues and the quantitative corrections were not necessary.
TEST FOR COLOUR INTERFERENCE
The mixture of 30 µL of the test item per 300 µl aqua dest. showed no colouring detectable by unaided eye-assessment. The mixture of 30 µL of the test item per 300 µl isopropanol showed colouring detectable by unaided eye-assessment. Therefore, the absorption of the chemical in isopropanol was measured in the range of 570 ± 30 nm. No absorption was measured in the relevant range. Therefore, an additional test with viable tissues (without MTT addition) was not necessary (NSC equalled 0%).
ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: mean absolute OD570 of the three negative control tissues was ≥ 0.8 and ≤ 2.8 (value: 1.849).
- Acceptance criteria met for positive control: mean relative tissue viability (% negative control) of the positive control was ≤ 20% (3.2 %)
- Acceptance criteria met for variability between replicate measurements: standard deviation of viability of replicate tissues of all dose groups was ≤ 18% (0.2 % - 2.4 %).
- The absorbance values were not below historically established boundaries.
Please also refer to the field "An other information on results incl. tables" below. - Interpretation of results:
- other: Category 1 (corrosive) or Category 2 (irritant) based on GHS criteria
- Conclusions:
- The test item, bismuth tris(2-ethylhexanoate), is either corrosive or irritating to the skin. Since the RhE test methods covered by OECD TG 439 cannot resolve between UN GHS Categories 1 or 2, further information on skin corrosion is required to decide on its final classification.
Referenceopen allclose all
Results of the test item bismuth tris (2-ethylhexanoate)
|
CORROSITEX™ Time [min] |
Colour Change |
Consistency Change |
Replicate 1 |
> 60 |
no |
no |
Replicate 2 |
> 60 |
no |
no |
Replicate 3 |
> 60 |
no |
no |
Replicate 4 |
> 60 |
no |
no |
Mean ± SD |
> 60 |
|
|
|
|
|
|
Positive control |
26.18 |
yes |
no |
Negative control |
> 60 |
no |
no |
The mean time, required to activate the CDS was > 60min.
Historical Data
Mean time [min] until colour change of positive control (Phosphoric Acid |
Standard Deviation [min] |
number of control values |
Range of LCL - UCL [min] |
21.4 |
13.3 |
35 |
0.0 – 48.0 |
LCL: Lower control limit (95%, mean – 2*SD)
UCL: Upper control limit (95%, mean + 2*SD)
Historical data were generated from 2010 - 2018
Table1: Result of the test item bismuth tris (2-ethylhexanoate)
Name |
Negative Control |
Positive Control |
Test Item |
||||||
Tissue |
1 |
2 |
3 |
1 |
2 |
3 |
1 |
2 |
3 |
Absolute OD570 |
1.824 |
1.879 |
1.802 |
0.090 |
0.106 |
0.099 |
0.130 |
0.124 |
0.128 |
1.898 |
1.893 |
1.800 |
0.094 |
0.105 |
0.109 |
0.133 |
0.128 |
0.133 |
|
Mean Absolute OD570 |
1.849**** |
0.101 |
0.129 |
||||||
OD570(Blank Corrected) |
1.781 |
1.836 |
1.759 |
0.047 |
0.063 |
0.056 |
0.087 |
0.080 |
0.085 |
1.855 |
1.849 |
1.757 |
0.051 |
0.062 |
0.066 |
0.090 |
0.085 |
0.090 |
|
Mean OD570of the Duplicates (Blank Corrected) |
1.818 |
1.843 |
1.758 |
0.049 |
0.063 |
0.061 |
0.088 |
0.083 |
0.087 |
Total Mean OD570of 3 Replicate Tissues (Blank Corrected) |
1.806* |
0.058 |
0.086 |
||||||
SD OD570 |
0.044 |
0.007 |
0.003 |
||||||
Relative Tissue Viability [%] |
100.6 |
102.0 |
97.3 |
2.7 |
3.5 |
3.4 |
4.9 |
4.6 |
4.8 |
Mean Relative Tissue Viability [%] |
100.0 |
3.2** |
4.8 |
||||||
SD Tissue Viability [%] |
2.4*** |
0.4*** |
0.2*** |
||||||
CV [% Viabilities] |
2.4 |
12.9 |
3.4 |
*Blank-corrected mean OD570 nmof the negative control corresponds to 100% absolute tissue viability.
**Mean relative tissue viability of the three positive control tissues is ≤ 20%.
***Standard deviation (SD) obtained from the three concurrently tested tissues is≤ 18%
****Mean absolute OD570of the negative control tissues is≥0.8 and≤2.8.
Table2: Historical Data
|
Mean Absolute OD570±30nmNC |
MeanAbsoluteOD570±30nmPC |
Mean Relative Viability [%] PC |
SD Viability [%] |
Mean |
1.861 |
0.114 |
3.7 |
4.4 |
SD |
0.247 |
0.033 |
1.5 |
4.1 |
Range of |
1.367 – 2.355 |
0.048 – 0.181 |
0.7 – 6.8 |
0.0 – 12.5 |
n |
25 |
25 |
25 |
117 |
LCL: Lower control limit (95%, mean – 2*SD)
UCL: Upper control limit (95%, mean + 2*SD)
n: number of control values
Historical data were generated in 2017.
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irritating)
Eye irritation
Link to relevant study records
- Endpoint:
- eye irritation: in vivo
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 2018-09-24 to 2018-10-16
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 405 (Acute Eye Irritation / Corrosion)
- Version / remarks:
- 2017-10-09
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- signed on 2018-04-26
- Species:
- rabbit
- Strain:
- New Zealand White
- Details on test animals or tissues and environmental conditions:
-
TEST ANIMALS
- Source: Charles River Deutschland, 97633 Sulzfeld, Germany
- Age at study initiation: approximately 12 – 13 weeks old
- Weight at study initiation: 2.3 kg
- Housing: individually housed in ABS-plastic or Noryl rabbit cages, floor 4200 cm²
- Diet (ad libitum): autoclaved hay and to Altromin 2123 maintenance diet for rabbits, rich in crude fibre
- Water (ad libitum): tap water
- Acclimation period: at least 5 days
ENVIRONMENTAL CONDITIONS
- Temperature: 18 ± 3 °C
- Relative humidity: 55 ± 10%
- Air changes: at least 10 x / hour
- Photoperiod (hrs dark / hrs light): 12/12 - Vehicle:
- unchanged (no vehicle)
- Controls:
- no
- Amount / concentration applied:
- TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 ml of the test item applied in the conjunctival sac of one eye. Untreated eye served as control. - Duration of treatment / exposure:
- not applicable
- Observation period (in vivo):
- 1, 24, 48 and 72 hours as well as 4 to 21 days after test item application
- Duration of post- treatment incubation (in vitro):
- not applicable
- Number of animals or in vitro replicates:
- 1 male rabbit
- Details on study design:
- PREPARATION OF THE ANIMALS
Within 24 hours before the test and immediately prior to the application both eyes of the animal were examined.
Approx. 15 hours before the application the eyes were examined with the aid of a fluorescein solution (Fluoreszein SE Thilo®). The eyes were rinsed with physiological saline 0.9 % NaCl after the examination. The animal did not show eye irritation, ocular defects, or pre-existing corneal injury.
INITIAL AND CONFIRMATORY TEST
The in vivo test was performed initially using one animal. The results of the initial test indicated that the test item is corrosive or severe irritant to the eye using the procedure described. Therefore, no additional animals were treated due to animal welfare reasons.
USE OF TOPICAL ANESTHETICS AND SYSTEMIC ANALGESICS
One hour before the application of the test item, 0.01 mg/kg of buprenorphine (Temgesic®) was administered subcutaneously in order to achieve a therapeutic level of systemic analgesia.
Approx. 5 minutes prior to the application of the test item, 1-2 drops of an ocular anaesthetic (Proparakaine-POS® hydrochloride ophthalmic 0.5% solution) were administered in both the treated and the control eye of the animal.
To prevent pain and distress after the application of the test item the animal was treated with doses of buprenorphine and meloxicam to provide a continued therapeutic level of systemic analgesia:
- After test substance application the test animal was dosed with 0.03 mg/kg bw of buprenorphine at the discretion of the veterinarian: 11 hours (day 0) post-application once; day 1 and day 2 twice; day 3 once.
- After test substance application the test animal was dosed with 0.5 mg/kg bw of meloxicam: 8 hours (day 0) up to day 5 and from day 7 up to day 21.
- Day 6, 20 and 21 no analgesic medication.
REMOVAL OF TEST SUBSTANCE
- Washing: treated eye was rinsed with physiological saline 0.9 % NaCl after examination with fluorescein solution
- Time after start of exposure: 1 hour after the application
The eyes were not rinsed to remove residues of the test item, foreign bodies or incrustation 24 hours after application.
SCORING SYSTEM
according to Draize scale
TOOL USED TO ASSESS SCORE: slit lamp biomicroscope / fluorescein
Approximately 15 hours before the application the eyes were examined with the aid of a fluorescein solution.
24 hours post-application and from then on daily until end of the observation period, the treated eye was examined with the aid of a fluorescein solution and a slit lamp biomicroscope. After the examination the eye was rinsed with physiological saline 0.9% NaCl.
OBSERVATIONS
- body weight: prior to the administration, 72 hours after administration and at the end of the observation period
- clinical observations: The eyes were comprehensively evaluated for the presence or absence of ocular lesions one hour and then 24, 48 and 72 hours after application. The eyes were further examined for signs of irritation throughout the observation period. - Irritation parameter:
- cornea opacity score
- Basis:
- mean
- Remarks:
- animal #1
- Time point:
- 24/48/72 h
- Score:
- 1
- Max. score:
- 4
- Reversibility:
- not reversible
- Irritation parameter:
- iris score
- Basis:
- mean
- Remarks:
- animal #1
- Time point:
- 24/48/72 h
- Score:
- 1.33
- Max. score:
- 2
- Reversibility:
- not reversible
- Irritation parameter:
- chemosis score
- Basis:
- mean
- Remarks:
- animal #1
- Time point:
- 24/48/72 h
- Score:
- 1.33
- Max. score:
- 4
- Reversibility:
- not reversible
- Irritation parameter:
- conjunctivae score
- Basis:
- mean
- Remarks:
- animal #1
- Time point:
- 24/48/72 h
- Score:
- 1.67
- Max. score:
- 3
- Reversibility:
- not reversible
- Irritant / corrosive response data:
- Animal # 1:
- conjunctival redness score 1 one hour post-application. 24 hours post-application a conjunctival redness score 2 was observed before the score decreased to 1 from 72 hours until 12 days post-application. No conjunctival redness was detected 13 days after installation of the test item. 14 to 21 days after dosing the animal showed conjunctival redness score 1. The effect was not fully reversible within the observation period of 21 days after application of the test material. The mean score of animal no. 1, following grading at 24, 48 and 72 h after installation of Bismuth tris (2-ethylhexanoate), was calculated to be 1.66.
- conjunctival chemosis score 3 one hour and score 2 24 hours post-application. From 72 hours to 11 days post-administration of the test material conjunctival chemosis was assessed with score 1. 12 days after test item installation score 2 was observed, before the score decreased again to 1 from 13 days post-application until end of the observation period. The effect was not fully reversible within the 21 days-observation period. The mean score of animal no. 1, following grading at 24, 48 and 72 h after installation of Bismuth tris (2-ethylhexanoate), was calculated to be 1.33.
- Iris lesion score 1 one hour post-application and score 2 24 hours post-application. From 72 hours until end of the observation period iris lesion score 1 was observed in the tested animal. The effect was not fully reversible within the observation period of 21 days after dosing. The mean score of animal no. 1, following grading at 24, 48 and 72 h after installation of Bismuth tris (2-ethylhexanoate), was calculated to be 1.33.
- Cornea opacity score 1 throughout the 21 days-observation period. The effect was not fully reversible within the test period. The mean score of animal no.1, following grading at 24, 48 and 72 h after installation of the test material, was calculated to be 1.00.
Local effects were observed in animal #1:
The observed local effects comprised slight to moderate hypersecretion, half eyelid closure and incline of the head.
Upon fluorescein examinations starting 24 hours post-application, the treated eyes of animal no. 1 showed corneal daily lesions (starting with approx. 100% of the area) until 12 days post-application. From 13 days to 19 days after administration of the test item no stainable lesions were recorded in the test animal, while from 20 days to 21 days post-application slightly visible changes in the cornea were apparent (approx. 10% of the area). - Other effects:
- - clinical observations: neither mortality nor significant clinical signs of toxicity were observed.
- body weight: the body weight development was within the expected range. - Interpretation of results:
- Category 1 (irreversible effects on the eye) based on GHS criteria
- Conclusions:
- The substance is serious eye damaging.
According to Annex I of Regulation (EC) 1272/2008 and subsequent adaptations, the test item Bismuth tris (2-ethylhexanoate) has to be classified as serious eye damaging (Category 1).
Reference
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed (irreversible damage)
Respiratory irritation
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
Skin corrosion/irritation:
The substance was observed to be either corrosive or irritating to the skin in a reliable in vitro skin irritation/corrosion study according to OECD 439. As the results of an OECD 439 test are not suitable to differentiate between skin categories 1 and 2, the substance was tested in an in vitro skin corrosion test.
Bismuth tris(2 -ethylhexanoate) was tested in an in vitro skin corrosion test according to OECD 435 and the substance was not observed to be corrosive to the skin.
Eye irritation:
Bismuth tris(2 -ethylhexanoate) was tested in two in vitro test systems evaluating eye damaging potential (OECD 437; GLP and OECD 492; GLP) and in an in vivo test (OECD 405).
The in vitro eye irritancy potential of Bismuth tris (2-ethylhexanoate) could not be assessed in the bovine corneal opacity and permeability assay, since the experiment showed residual of Bismuth tris (2-ethylhexanoate) on the corneas. The study could not be performed according to the guideline OECD 437.
With the OECD 492 study a prediction of the ocular irritation potential of Bismuth tris (2-ethylhexanoate) could not be made. In accordance with OECD 492 guideline further testing with other test methods will be required, because this test cannot resolve between UN GHS Categories 1 and 2.
Based on these results of both in-vitro tests further in-vivo testing was necessary for assessing the acute eye irritation/corrosion potential of the test substance. Bismuth tris (2-ethylhexanoate) produced irritant effects in the rabbit, which were not fully reversible within an observation period of 21 days.
Justification for classification or non-classification
Skin irritation:
Bismuth tris(2 -ethylhexanoate) does possess a skin irritation potential based on in vitro OECD 435 and 439 studies. The substance does require classification as skin irritant according to Regulation (EC) No 1272/2008 and its subsequent adaptations (Category 2; H315).
Eye irritation:
Bismuth tris(2 -ethylhexanoate) does possess serious eye damaging potential based on an in vivo OECD 405 study. The substance does require classification as serious eye damaging according to Regulation (EC) No 1272/2008 and its subsequent adaptations (Category 1; H318).
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.