lithium nickel dioxide

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

29 July 2016

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier): BASF SE, Batch: 8800315
- Purity: approx. 99.8 g/100 g
- Expiration date of the batch: 12 Aug 2021

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: guaranteed by the sponsor

FORM AS APPLIED IN THE TEST: undiluted solid test substance

OTHER SPECIFICS:
- Pysical state/color: solid/anthracite
- pH: ca. 12 (undiluted test substance moistened with deionized water, determined in the lab prior to start of the GLP study)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Origin: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia. Tissue model: EPI-200

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm 200 kit, EPI-200

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

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: washing occurred after incubation
- Observable damage in the tissue due to washing: not described

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

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: two tissues in parallel, inter-tissue variability (range of 20% and 100% viability) accepted if the SD of % viability is ≤ 18%
- Negative and positive controls: OD570 of the negative control between 0.8 and 2.8 and of the positive control (8 N potassium hydroxide) ≤ 30 % for 3-min exposure; ≤ 15 % for 1-hour exposure
- Barrier function: lower acceptance limit: ET50 = 4.0 hours; upper acceptance limit: ET50 = 8.7 hours; both for Triton X-100 (1%)
Further information are given below in the section 'Any other information', table 1.

NUMBER OF REPLICATE TISSUES: 2

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- freeze-killed control tissues (KC)

PREDICTION MODEL / DECISION CRITERIA
Further details are given in table 2.
- The corrosive potential of the test materials is predicted from the mean relative tissue viabilities (3-minute treatment).
- The test substance is considered as "corrosive" if the mean relative tissue viability after the 3-minute treatment with a test material is < 50%
- The test substance is considered as "corrosive" if viability of ≥ 50% after the 3-minute treatment and the mean relative tissue viability after a 1-hour treatment is < 15%.
- In case of borderline results such as non-concordant replicate measurements and/or mean percent tissue viability equal to ± 5% of the cutoff value, a second test (consisting of two tissue replicates) should be considered as well as a third one in case of discordant results between the first two tests.

- Justification for the selection of the cut-off point:
- A “borderline“ range (50 ± 5%, 25 ± 5% and 15 ± 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 431.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- Amount applied: a bulk volume of ca. 25 μL 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
- Concentration: 8 N

Duration of treatment / exposure:

3 minutes (room temperature) or
1 hour (incuibator at 37°C)

Number of replicates:

2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean value from two tissues, 3 min exposure

Value:

7.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: corrosive

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean value from three tissues, 1 h exposure

Value:

3.2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: corrosive

Other effects / acceptance of results:

- Direct-MTT reduction:
Due to the ability of the test substance to reduce MTT directly, KC tissues were applied in parallel. However, the results of the KC tissues indicated no increased MTT reduction and no interference due to the colored compound residues.

DEMONSTRATION OF TECHNICAL PROFICIENCY: based on the historical data, a profound experience with the assay is assumed

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for positive control: The mean OD570 of the negative control (deionized water) fulfills the acceptance criteria and demonstrates the validity of the assay.
- Acceptance criteria met for negative control: The mean OD570 of the negative control (deionized water) fulfills the acceptance criteria and demonstrates the validity of the assay.

Moderate black colored compound residues remained on the tissues treated with the test substance after the washing procedure.

 

Table 3: Individual and mean OD570 values, individual and mean viability values, standard deviations and coefficient of variation after 3 min exposure

Exposure period: 3 min
Test substance identification			tissue 1	tissue 2	mean	SD	CV [%]
NC	viable tissues	mean OD570	1.659	1.758	1.709
		viability [% of NC]	97.1	102.9	100.0	4.1	4.1
	KC tissues	mean OD570	0.077	0.070	0.074
		viability [% of NC]	4.5	4.1	4.03	0.3	7.2
test substance	viable tissues	mean OD570	0.133	0.146	0.130
		viability [% of NC]	6.6	8.6	7.6	1.4	18.3
	KC tissues*	mean OD570 KC NC corrected	0.000	0.000	0.000
		viability [% of NC]	0.0	0.0	0.0
PC	viable tissues	mean OD570	0.187	0.158	0.172
		viability [% of NC]	10.9	9.2	10.1	1.2	11.9

* Negative values are set to zero for further calculation

 

Table 4: Individual and mean OD570 values, individual and mean viability values, standard deviations and coefficient of variation after 1 hour exposure

 

Exposure period: 1 h
Test substance identification			tissue 1	tissue 2	mean	SD	CV [%]
NC	viable tissues	mean OD570	1.649	1.785	1.717
		viability [% of NC]	96.0	104.0	100.0	5.6	5.6
	KC tissues	mean OD570	0.060	0.060	0.060
		viability [% of NC]	3.5	3.5	3.5	0.0	0.6
test substance	viable tissues	mean OD570	0.055	0.054	0.054
		viability [% of NC]	3.2	3.1	3.2	0.1	2.0
	KC tissues*	mean OD570 KC NC corrected	0.000	0.000	0.000
		viability [% of NC]	0.0	0.0	0.0
PC	viable tissues	mean OD570	0.086	0.099	0.092
		viability [% of NC]	5.0	5.7	5.4	0.5	9.6

* Negative values are set to zero for further calculation

Cross-reference 2

Reason / purpose for cross-reference:

reference to same study

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Reason / purpose for cross-reference:

reference to same study

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

26 June 2020

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier): BASF SE, Batch: 8800315
- Purity: approx. 99.8 g/100 g
- Expiration date of the batch: 12 Aug 2021

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: guaranteed by the sponsor

FORM AS APPLIED IN THE TEST: undiluted solid test substance

OTHER SPECIFICS:
- Pysical state/color: solid/anthracite
- pH: ca. 12 (undiluted test substance moistened with deionized water, determined in the lab prior to start of the GLP study)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Origin: MatTek In Vitro Life Science Laboratories, Bratislava, Slovakia. Tissue model: EPI-200

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used:
EpiDerm 200 kit, EPI-200

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

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps:
washing occurred after incubation and post-incubation
- Observable damage in the tissue due to washing:
not described

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

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: three tissues in parallel, inter-tissue variability accepted if the SD of % viability is ≤ 18%
- Negative and positive controls: OD570 of the negative control between 0.8 and 2.8 and of the positive control (5% SDS) ≤ 20%
- Barrier function: lower acceptance limit: ET50 = 4.0 hours; upper acceptance limit: ET50 = 8.7 hours; both for Triton X-100 (1%)
Further information are given below in the section 'Any other information'.

NUMBER OF REPLICATE TISSUES:
3

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- freeze-killed control tissues (KC)

PREDICTION MODEL / DECISION CRITERIA
- The test substance is identified as requiring classification and labelling according to UN GHS (Category 2 or Category 1) if the mean tissue viability after exposure and post-treatment incubation is less than or equal (≤) to 50%.
- The test substance is considered to be non-irritant to skin if the viability after exposure is greater than 50 %.
- In case of borderline results such as non-concordant replicate measurements and/or mean percent tissue viability equal to ± 5% of the cutoff value, a second test (consisting of three tissue replicates) should be considered as well as a third one in case of discordant results between the first two tests.
- Justification for the selection of the cut-off point:
- A “borderline“ range (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

Amount/concentration applied:

TEST MATERIAL
- Amount applied: a bulk volume of ca. 25 μL 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
- Concentration: 5% (w/v)

Duration of treatment / exposure:

25 minutes at room temperature and for 35 minutes in the incubator at 37°C

Duration of post-treatment incubation (if applicable):

24 ± 2 hour

Number of replicates:

3

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

mean value from three tissues out of one experiment

Value:

2.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: No prediction can be made (UN GHS Category 2 or 1)

Other effects / acceptance of results:

- Direct-MTT reduction:
Due to the ability of the test substance to reduce MTT directly, KC tissues were applied in parallel. However, the results of the KC tissues indicated no increased MTT reduction and no interference due to the colored compound residues.

DEMONSTRATION OF TECHNICAL PROFICIENCY:
based on the historical data, a profound experience with the assay is assumed


ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: The mean OD570 of the negative control (PBS) fulfills the acceptance criteria and demonstrates the validity of the assay.
- Acceptance criteria met for positive control: The tissues treated with the positive control (5% SDS) showed a relative mean viability of 2.6%, reflecting the expected sensitivity of the tissues.

Table 2: Individual and mean OD570 values, individual and mean viability values, standard deviations and coefficient of variation

Test substance identification			tissue 1	tissue 2	tissue 3	mean	SD	CV [%]
NC	viable tissues	mean OD570	1.639	1.826	1.660	1.708
		viability [% of NC]	96.0	106.9	97.2	100.0	6.0	6.0
	KC tissues	mean OD570	0.040	0.043	0.039	0.041
		viability [% of NC]	2.4	2.5	2.3	2.4	0.1	4.9
test substance	viable tissues	mean OD570	0.046	0.046	0.050	0.047
		viability [% of NC]	2.7	2.7	2.9	2.7	0.1	4.9
	KC tissues*	mean OD570 KC NC corrected	0.000	0.000	0.000	0.000
		viability [% of NC]	0.0	0.0	0.0	0.0
PC	viable tissues	mean OD570	0.048	0.042	0.46	0.045
		viability [% of NC]	2.8	2.4	2.7	2.6	0.2	7.3

* Negative values are set to zero for further calculation

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2021

Report date:

2021

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)

Version / remarks:

28 July 2015

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Lithium; oxido(oxo)nickel

Cas Number:

12031-65-1

Molecular formula:

LiNiO2

IUPAC Name:

Lithium; oxido(oxo)nickel

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source (i.e. manufacturer or supplier): BASF SE, Batch: 8800315
- Purity: approx. 99.8 g/100 g
- Expiration date of the batch: 12 Aug 2021

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature
- Stability under test conditions: guaranteed by the sponsor

FORM AS APPLIED IN THE TEST: undiluted solid test substance

OTHER SPECIFICS:
- Pysical state/color: solid/anthracite
- pH: ca. 12 (undiluted test substance moistened with deionized water, determined in the lab prior to start of the GLP study)

In vitro test system

Test system:

artificial membrane barrier model

Vehicle:

unchanged (no vehicle)

Details on test system:

MATERIAL:

Corrositex® kit:
InVitro International, Irvine CA, USA, containing: reagents required for qualification and categorization screen, biobarrier matrix powder and diluent, membrane discs and vials containing the Chemical Detection System.

Controls:
Negative control (NC): 10% citric acid
Positive control (PC): Sodium hydroxide (solid)

EXPERIMENTAL PROCEDURE:
- Test substance compatibility with the assay (qualification screen):
For the qualification screen, 100 mg test substance were added to the CDS screening tube. If the test substance failed to produce a color change in the CDS within one minute, the test substance could not be analyzed in this system, and no further testing was required.
- Categorization screen:
The categorization screen was used to assess the appropriate scoring scale for the test substance. The categorization screen was performed by adding 100 mg test substance to tube A and B each. Each tube was mixed, and the resulting color was observed. If required, 2 drops of the "confirm" reagent were added to tube B, the tube was mixed and the resulting color observed. The categorization kit and color chart provided by InVitro International were used to determine
the category. The test substance was scored as category 1 (high acid/alkaline reserve) or category 2 (low acid/alkaline reserve) as described in section 3.9.3.
- Bio barrier preparation:
The entire content of the biobarrier diluent vial was added slowly to the vial containing the bio barrier matrix powder. The vial containing the mixture was placed in a water bath at 64 – 68°C with a magnetic stirrer. The stir bar rotated slowly enough to avoid foaming until complete and uniform solubilisation. 200 μL solubilized matrix were pipetted into each of the membrane discs.
The membrane discs were then refrigerated for at least 2 hours at 2 – 8°C.
The bio barriers were wrapped and stored at 2 – 8°C for a maximum of 7 days.
Any remaining matrix solution was stored at 2 – 8°C for up to 30 days for preparation of additional bio barrier membrane discs.
- Corrositex® assay:
Following the acceptance of the positive control, the Corrositex® assay was performed for the test substance. Four vials containing the CDS were used for the test substance. In addition, one vial was used for the PC, the NC and the color control (blank) each. A membrane disc coated with the bio barrier matrix was placed into one vial containing the CDS, and approximately 500 mg test substance were added onto the membrane disc. An electronic time clock was started with the application. The vial was observed for three minutes for any change in the CDS. If no color change was observed within three minutes, the membranes remaining were treated with the test substance. An electronic time clock was started with each application. The vials were observed continuously for the first ten minutes. Thereafter, the vials were observed for
approximately ten minutes around the time points relevant for evaluation or until breakthrough of the test substance. The elapsed time between test substance
application and the first change in the indicator solution (i.e. barrier penetration) was recorded.
The positive control vial was prepared as described above and contained one pellet of sodium hydroxide on top of the membrane disc. This vial was monitored continuously until breakthrough.
The negative control vial was prepared as described above and contained 500 μL 10% citric acid monohydrate. This vial was observed for 60 minutes and evaluated as “non-corrosive” if no reaction had been observed.

ACCEPTANCE CRITERIA:
The Corrositex® assay was accepted if the breakthrough time for the positive control substance was in the historic control range (mean ± 2-3x standard deviations). The expected breakthrough time of the concurrent positive control (Sodium Hydroxide solid) should be between 8 - 16 min. In order to demonstrate the functional integrity of the membrane barrier, the acceptance criterium for the negative control was not to induce membrance breakthrough within a 60- minute observation period.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

500 mg test substance

Duration of treatment / exposure:

max. 3 minutes

Number of replicates:

4

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: mean breakthrough time through Corrositex® Biobarrier Membrane in minutes

Run / experiment:

Mean of 4 vials

Value:

58.39

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: corrosive subcategory 1C

Any other information on results incl. tables

The qualification screen demonstrated that the test substance is able to react with the CDS and produce a visible color change. Therefore, the membrane barrier test method was determined to be suitable for the evaluation of the corrosive potential of the test substance.
A timescale category test was carried out to distinguish between weak and strong acids or bases. The test substance was assigned to timescale category 2 (having a low acid/alkaline reserve).
The mean breakthrough time of the test substance determined in the actual Corrositex® assay was 58 minutes and 39 seconds (single break through times of the four vials [min:s]: 59:59, 67:301, 59:22 and 56:37). The second vial treated with the test substance showed a breakthrough time above 60 minutes, indicating no corrosive potential. However, break through times below 60 minutes were noted in the other three vials, thus, the test material is classified as corrosive.
The breakthrough times of three vials, indicated that the test substance has a weak corrosive potential and should be assigned to UN GHS skin corrosivity subcategories 1C or UN Transport Packing Group III as specified in the cited OECD TG 435 and Instruction Manual.

The negative control, 10% citric acid monohydrate, did not produce any reaction within 60 minutes after application. Sodium hydroxide (solid) applied as positive control showed a breakthrough time of 15 minutes and 55 seconds and was assigned accordingly. Thus, the controls fulfill the acceptance criteria and demonstrate the validity of the assay.

Applicant's summary and conclusion

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Remarks:

Migrated information Criteria used for interpretation of results: EU