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Administrative data

Description of key information

Skin irritation/corrosion: Based on an in vitro test battery and an in vivo study in rabbits the substance shows no skin irritation potential.

Eye irritation: Based on an in vivo study the test substance was considered to cause serious eye damage in rabbits.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Referenceopen allclose all

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
26 July 2016 - 16 September 2016
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:
28 July 2015
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EU Method B.40 BIS.: Methods for the determination of toxicity and other health effects: In Vitro Skin Corrosion: Human Skin Model Test; Official Journal of the European Union, No. L 142
Version / remarks:
30 May 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: (IATA) for skin corrosion and irritation, Series on Testing and Assessment No. 203
Version / remarks:
11 July 2014
GLP compliance:
yes (incl. QA statement)
Remarks:
BASF SE Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Do_0164

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, under nitrogen
- Stability under test conditions: guaranteed
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ 200 kit ( EPI-200 tissues with surface of 0.6 cm²), MatTek In Vitro LifeScience Laboratories, Bratislava, Slovakia
- Tissue batch number(s): 23348

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 3 min at room temperature or 1 hour in the incubator at 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing: washed with PBS to remove residual test material

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg / mL
- Incubation time: 3 hours
- Spectrophotometer: SunriseTM Absorbance Reader
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: OD (540-570) 1.850 ± 0.053
- Barrier function:ET50 = 6.18 hours
(Lower acceptance limit: ET50 = 4.0 hours and Upper acceptance limit: ET50 = 8.7 hours)
- Morphology: functional stratum corneum, a viable basal cell layer, intermediate spinous and granular layers
- Contamination: no, sterile

NUMBER OF REPLICATE TISSUES: 2 tissues per exposure time

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- killed tissues
- Procedure used to prepare the killed tissues: EPI-200 tissue that is killed by freezing at -20°C
- N. of replicates : 2

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

PREDICTION MODEL / DECISION CRITERIA
Mean tissue viability
(% of negative control)
3 min: < 45 corrosive (Optional Sub-category 1A) (1)
3 min: 45-55 borderline (2)
3 min: >55 and 1 hour: 10-20 borderline (2)
3 min: >55 and 1 hour: < 10 Corrosive (Optional Sub-category 1B and 1C) (1)
3 min: >55 and 1 hour: >20 Non-corrosive
(1)According to the current OECD Guideline 431 a sub-categorization is possible based on the results. However, the sub-categorization into 1A is highly over-predictive as stated in the guideline and differentiation into sub-category 1B or 1C is not possible. If the test substance is identified to be corrosive by SCT and a transport classification is needed, the Corrositex® test should be performed, if applicable, to confirm classification as 1A or to differentiate between 1B and 1C.
(2) The „borderline“-evaluation (50 ± 5% and 15 ± 5%) was determined statistically using historic BASF data and hence considers the variance of the test method. This evaluation is an amendment to the evaluation provided in OECD Guideline 431.




Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 25 μL de-ionized water was applied first. Thereafter, a bulk volume of ca. 25 μL of the solid test material was applied with a sharp spoon and homogeneously distributed with the water.

NEGATIVE CONTROL
- Amount applied: 50 μL

POSITIVE CONTROL
- Amount applied: 50 μL
Duration of treatment / exposure:
3 min or 1 hour
Number of replicates:
2
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1st Experiment (3 min)
Value:
74.7
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
14.2%
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1st Experiment (1 hour)
Value:
38.6
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
5.5%
Other effects / acceptance of results:
- OTHER EFFECTS:
- Direct-MTT reduction: yes

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
Tissue viability is acceptable if the mean OD570 of the NC is ≥ 0.8. The mean OD570 of the NC should not exceed 2.8.
- Acceptance criteria met for positive control: yes
A tissue viability of ≤ 30% is acceptable for the 3-minute exposure. Mean viability of the tissues exposed for 1 hour should be <15%.
- Acceptance criteria met for variability between replicate measurements: no
In the range of 20% and 100% viability, variability between the tissues is considered to be acceptable if the coefficient of variation (CV) of %-viability is ≤ 30%. The CV of %-viability of the test substance is out of the acceptance range (46% , 1 hour exposure only). Since all other quality criteria of the test were met and the viability values are well above the cut off for skin corrosion, this deviation is not considered to adversely affect the result of this study.

Historic control data:
Historic Range of NC (Period Jan 2014 - Jun 2016)
3 min exposure: Mean OD570 1.945 ± 0.169
60 min exposure: Mean OD570 1.936 ± 0.172
Historic Range of PC (Period Jan 2014 - Jun 2016)
3 min exposure: Mean OD570 0.312 ± 0.074
60 min exposure: Mean OD570 0.130 ± 0.036
Viability % (Period Jan 2014 - Jun 2016)
3 min exposure: Mean % 16.1 ± 3.8
60 min exposure: Mean % 6.6 ± 1.7
Interpretation of results:
GHS criteria not met
Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
26 July 2016 - 16 September 2016
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
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
6 July 2012
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: (IATA) for skin corrosion and irritation, Series on Testing and Assessment No. 203
Version / remarks:
11 July 2014
GLP compliance:
yes (incl. QA statement)
Remarks:
BASF SE Experimental Toxicology and Ecology, 67056 Ludwigshafen, Germany
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Do_0164

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, under nitrogen
- Stability under test conditions: guaranteed
Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ 200 kit ( EPI-200 tissues with surface of 0.6 cm²), MatTek In Vitro LifeScience Laboratories, Bratislava, Slovakia
- Tissue batch number(s): 23348

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature for 25 minutes overall and for 35 minutes in the incubator at 37°C
- Temperature of post-treatment incubation: 37°C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Washing: with sterile PBS

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg / mL
- Incubation time: 3 hours
- Spectrophotometer: SunriseTM Absorbance Reader
- Wavelength: 570 nm

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: OD (540-570) 1.850 ± 0.053
- Barrier function:ET50 = 6.18 hours
(Lower acceptance limit: ET50 = 4.0 hours and Upper acceptance limit: ET50 = 8.7 hours)
- Morphology: functional stratum corneum, a viable basal cell layer, intermediate spinous and granular layers
- Contamination: no, sterile

NUMBER OF REPLICATE TISSUES: 3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- killed tissues
- Procedure used to prepare the killed tissues: EPI-200 tissue that is killed by freezing at -20°C
- N. of replicates : 3

PREDICTION MODEL / DECISION CRITERIA
Mean tissue viability
(% of negative control) Prediction
< 45 Irritant
45 - 55 Borderline (1)
> 55 Non-irritant
(1)The „borderline“-evaluation (50 ± 5%) was determined statistically using historic BASF data and hence considers the variance of the test method. This evaluation is confirming the borderline range provided in OECD Guideline 439.


Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
TEST MATERIAL
- Amount applied: 25 μL sterile PBS was applied first. Thereafter, a bulk volume of ca. 25 μL of the solid test material was applied with a sharp spoon and homogeneously distributed together with the fluid.

NEGATIVE CONTROL
- Amount applied: 30 μL

POSITIVE CONTROL
- Amount applied: 30 μL
Duration of treatment / exposure:
1 hour
Duration of post-treatment incubation (if applicable):
ca. 42 hours
Number of replicates:
3
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1st Experiment
Value:
85.7
Negative controls validity:
valid
Remarks:
100%
Positive controls validity:
valid
Remarks:
3.3%
Other effects / acceptance of results:
- OTHER EFFECTS:
- Direct-MTT reduction: yes

DEMONSTRATION OF TECHNICAL PROFICIENCY: yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
Tissue viability is acceptable if the mean OD570 of the NC is ≥ 0.8. The mean OD570 of the NC should not exceed 2.8.
- Acceptance criteria met for positive control: yes
A viability of ≤ 20% is acceptable.
- Acceptance criteria met for variability between replicate measurements: yes
In the range of 20% and 100% viability, variability between the tissues is considered to be acceptable if the coefficient of variation (CV) of %-viability is ≤ 30%.

Historic control data:
Historic Range of NC (Period Jan 2014 - Jun 2016)
Mean OD570 2.324 ± 0.276
Historic Range of PC (Period Jan 2014 - Jun 2016)
Mean OD570 0.071 ± 0.011
Viability % (Period Jan 2014 - Jun 2016)
Mean % 3.1 ± 0.5
Interpretation of results:
GHS criteria not met
Endpoint:
skin irritation: in vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
test procedure in accordance with national standard methods with acceptable restrictions
Qualifier:
according to guideline
Guideline:
other: Draize test according to Federal Register 38, No. 187, § 1500.41, p. 27019, 27.09.73.
Deviations:
no
GLP compliance:
no
Species:
rabbit
Strain:
Vienna White
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Diet: The animals were offered a standardized animal laboratory diet, ad libitum
Type of coverage:
occlusive
Preparation of test site:
other: abraded and intact skin
Vehicle:
water
Controls:
other: untreated skin of the same animal
Amount / concentration applied:
50 µL of an 50% aqueous preparation
Duration of treatment / exposure:
24 hours
Observation period:
8 days
Number of animals:
3
Details on study design:
TEST SITE
- Area of exposure: back
- % coverage: 2.5 x 2.5 cm

REMOVAL OF TEST SUBSTANCE
- Washing: no

READINGS: 24 h, 72 h, 8 d

SCORING SYSTEM: erythema and edema according to Draize (1959)
No symptom: 0
Questionable (redness)/very slight (edema): 1
Slight: 2
Marked: 3
Severe: 4
Irritation parameter:
erythema score
Remarks:
intact skin
Basis:
animal #1
Time point:
other: 24, 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Remarks:
intact skin
Basis:
animal #2
Time point:
other: 24, 72 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Remarks:
intact skin
Basis:
animal #3
Time point:
other: 24 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
erythema score
Remarks:
intact skin
Basis:
animal #3
Time point:
other: 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
animal #1
Time point:
other: 24, 72 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
animal #2
Time point:
other: 24 h
Score:
2
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
animal #2
Time point:
other: 72 h
Score:
0
Max. score:
4
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
animal #3
Time point:
other: 24 h
Score:
1
Max. score:
4
Reversibility:
fully reversible within: 8 days
Irritation parameter:
edema score
Remarks:
intact skin
Basis:
animal #3
Time point:
other: 72 h
Score:
0
Max. score:
4
Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records
Reference
Endpoint:
eye irritation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Qualifier:
according to guideline
Guideline:
other: Draize test according to Federal Register 38, No. 187, § 1500.42, p. 27019, 27.09.73.
Deviations:
no
GLP compliance:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 78/574
Species:
rabbit
Strain:
Vienna White
Details on test animals or tissues and environmental conditions:
TEST ANIMALS
- Diet: The animals were offered a standardized animal laboratory diet, ad libitum
Vehicle:
water
Controls:
other: untreated eye of the same animal
Amount / concentration applied:
50 µL of an 80% aqueous preparation
Duration of treatment / exposure:
24 hours
Observation period (in vivo):
8 days
Number of animals or in vitro replicates:
3
Details on study design:
APPLICATION
- Application site: single application to the conjunctival sac

REMOVAL OF TEST SUBSTANCE
- Washing: the test substance was not washed out

READINGS: 24 h, 48 h, 72 h, 8 d

SCORING SYSTEM: According to Draize (1959)
Cornea:
No symptom: 0
Slight: 1
Marked: 2
Severe: 3
Very severe: 4
Iris:
No symptom: 0
ciliar injection: 1
iritis: 2
Conjunctivae:
No symptom: 0
Slight: 1
Marked: 2
Severe: 3


Irritation parameter:
cornea opacity score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
cornea opacity score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0.67
Max. score:
4
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
cornea opacity score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
4
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
iris score
Basis:
animal #1
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 8 days
Remarks on result:
other: ingrowing vessels
Irritation parameter:
iris score
Basis:
animal #2
Time point:
24/48/72 h
Score:
0
Max. score:
2
Irritation parameter:
iris score
Basis:
animal #3
Time point:
24/48/72 h
Score:
1
Max. score:
2
Reversibility:
fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Basis:
animal #2
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
conjunctivae score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Irritation parameter:
chemosis score
Basis:
animal #1
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Remarks on result:
other: at day 8: score 2, ulceration, scar
Irritation parameter:
chemosis score
Basis:
animal #2
Time point:
24/48/72 h
Score:
1.33
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Remarks on result:
other: at day 8: score 1, scar, blood
Irritation parameter:
chemosis score
Basis:
animal #3
Time point:
24/48/72 h
Score:
2
Max. score:
3
Reversibility:
not fully reversible within: 8 days
Remarks on result:
other: at day 8: score 2, scar, ulceration
Interpretation of results:
Category 1 (irreversible effects on the eye) based on GHS criteria
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Skin irritation

The potential of the test item to cause dermal corrosion/irritation was assessed in two in vitro assays as part of an in vitro skin irritation and corrosion test strategy. Both tests were performed according OECD test guidelines (OECD TG 431 and OECD TG 439)

The test item was applied by a single topical application of ca. 25 μL bulk volume (about 14 mg) of the undiluted test substance to a reconstructed three dimensional human epidermis model (EpiDerm).

For the corrosion test two EpiDerm tissues were incubated with the test substance for 3 minutes and 1 hour, each. The irritation test was performed with three EpiDerm tissues, which were incubated with the test substance for 1 hour followed by a 42-hours post-incubation period.

Tissue destruction was determined by measuring the metabolic activity of the tissue after exposure/post-incubation using a colorimetric test. The reduction of mitochondrial dehydrogenase activity, measured by reduced formazan production after incubation with a tetrazolium salt (MTT) was chosen as endpoint. The formazan production of the test-substance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability.

The test substance is able to reduce MTT directly. Therefore an additional MTT reduction control KC (freeze-killed control tissues) was introduced.

Results of the Corrosion test:

The mean viability of the test-substance treated tissues determined after an exposure period of 3 minutes was 74.7%, and it was 38.6% after an exposure period of 1 hour. The variability of the test-substance treated tissues after the exposure period of 1 hour is out of the acceptance range. Since all other quality criteria of the test were met and the viability values are well above the cut off for skin corrosion, this deviation is not considered to adversely affect the result of this study.

Results of the Irritation test:

The mean viability of the test-substance treated tissues determined after an exposure period of 1 hour with about 42 hours post-incubation was 85.7%.

Based on the in vitro test strategy the substance shows no skin corrosion/irritation potential (BASF SE, 2017).

In addition, the skin irritation potential of the test substance was examined in a Draize test according to Federal Register 38, No. 187, § 1500.41 in rabbits. Three animals received a 50% aqueous solution of the test substance onto the intact and abraded skin for 24 hours under an occlusive dressing. The animals were observed over a period time of 8 days. The skin findings were recorded and scored at 24 and 72 hours according the scheme of Draize.

The mean erythema score for the intact skin was 2 at 24 hours and 1.7 at 72 hours, while the mean edema score for the intact skin was 1.3 at 24 hours and 0.3 at 72 hours. In any case, erythema and edema were completely reversible within 8 days. Based on these results the test substance was considered to be not irritating to the skin of rabbits (BASF SE, 1980).

Conclusion:

Based on the observed results in the in vitro test strategy and the in vivo study, the test item does not show a skin irritation potential under the test conditions chosen.

 

Eye irritation

The eye irritation potential of the test substance was examined in a Draize test according to Federal Register 38, No. 187, § 1500.42 in rabbits. Three animals received an 80% aqueous solution of the test substance into the conjunctival sac. The animals were observed over a period of 8 days. The eye findings were recorded and scored at 24, 48 and 72 hours according to the scheme of Draize. The mean cornea score after 24, 48 and 72 hours was determined to be 0.67 (1 animal) and 1 (two animals), which was not fully reversible within 8 days. The mean iris score after 24, 48 and 72 hours was 0 (1 animal) and 1 (2 animals), which was fully reversible within 8 days. The mean conjunctivae redness score after 24, 48 and 72 hours was 2 (3 animals), which was not fully reversible within 8 days. The mean chemosis score after 24, 48 and 72 hours was 1.33 (1 animal) and 2 (2 animals), which was not fully reversible within 8 days. Based on these results the test substance was considered to cause serious eye damage in rabbits.


Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. As a result the substance is not classified for skin irritation but considered to be classified for serious eye damage category 1 (H318: "Causes serious eye damage") under Regulation (EC) No 1272/2008, as amended for the tenth time in Regulation (EU) No 2017/776.