Morpholinium, 4-[2-[2-[(2-hydroxyphenyl)methylene]hydrazinyl]-2-oxoethyl]-4-methyl-, chloride (1:1)

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25.2.2011-08.09.2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 431, OECD 439

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

4-({N'-[(2-hydroxyphenyl)methylidene]hydrazinecarbonyl}methyl)-4-methylmorpholin-4-ium chloride

EC Number:

700-714-9

Cas Number:

1254469-57-2

Molecular formula:

C14H20ClN3O3

IUPAC Name:

4-({N'-[(2-hydroxyphenyl)methylidene]hydrazinecarbonyl}methyl)-4-methylmorpholin-4-ium chloride

Details on test material:

- Name of test material (as cited in study report): Tinocat ES 96000
- Analytical purity: The test substance has been characterized analytically (for details see report No.: 11L00011)
- Lot/batch No.: RD173323

- Stability under test conditions: The stability under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this
responsibility
- Other: pH-value: Approx. 4 (undiluted test substance, moistened with water)

Test animals

Species:

other: reconstructed human epidermal model

Details on test animals or test system and environmental conditions:

not applicable

Test system

Type of coverage:

other: not applicable

Preparation of test site:

other: not applicable

Vehicle:

other: water and PBS

Controls:

other: not applicable

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 25 μL bulk volume
- Concentration (if solution): about 11 mg

Duration of treatment / exposure:

Corrosion test: 3 minutes and 1 hour
Irritation test: 1 hour

Observation period:

Corrosion test: 0 hours
Irritation test: 42 hours

Number of animals:

Corrosion test: 2 tissues
Irritation test: 3 tissues

Details on study design:

not applicable

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

other: other: mean OD570 values (as measures of the viability) negative control

Value:

> 1 - < 2.5

Remarks on result:

other:

Remarks:

Basis: mean. Time point: 3 min or 1 hour. Remarks: tissue viability: quotient of the mean OD570 divided by the respective OD570 NC value in percent; Corrosion test: Acceptance criteria for PC (viability ≤ 30%) reached; Irritation test: Acceptance criteria for PC (viability ≤ 20%) reached. (migrated information)

Results 2

Irritation / corrosion parameter:

other: other: mean OD570 values (as measures of the viability) test item; corrosion test

Value:

92 - 98

Remarks on result:

other:

Remarks:

Basis: mean 2 tissues. Time point: 3 min or 1 hour. Remarks: Corrosion test: Mean tissue viability (% of negative control): 3 min: < 50 = corrosive; 3 min: ≥ 50 and 1 hour: < 15 = corrosive; 3 min: ≥ 50 and 1 hour: ≥ 15 = non-corrosive . (migrated information)

Results 3

Irritation / corrosion parameter:

other: other: mean OD570 values (as measures of the viability) test item; irritation test

Value:

32

Remarks on result:

other:

Remarks:

Basis: mean 3 tissues. Time point: 43 hours. Remarks: Irritation test: Mean tissue viability (% of negative control) ≤ 50 = irritant; > 50 = non-irritant . (migrated information)

Any other information on results incl. tables

Corrosion test data after 3 min

Test substance	Mean OD 570	Viability [% of negative control]
Negative control	1.804	100
11/0013-1	1.761	98
Positive control	0.368	20

Corrosion test data after 1 hour

Test substance	Mean OD 570	Viability [% of negative control]
Negative control	1.68	100
11/0013-1	1.543	92
Positive control	0.162	10

Irritation data

Test substance	Mean OD 570	Viability [% of negative control]
Negative control	1.890	100
11/0013-1	0.603	32
Positive control	0.123	7

Corrosive potential of the test materials is predicted from the mean relative tissue viabilities obtained after 3 min treatment compared to the negative control tissues concurrently treated with highly de-ionized water. A chemical is considered as "corrosive", if the mean relative tissue viability after 3 min treatment with a test material is decreased below 50%. In addition, those materials with a viability of ≥ 50% after 3 min treatment are considered as "corrosive" if the mean relative tissue viability after 1 hour treatment with a test material is decreased below 15%.

Irritant potential of the test materials is predicted from the mean relative tissue viabilities compared to the negative control tissues concurrently treated with sterile PBS. A chemical is considered as "irritant", if the mean relative tissue viability with a test material is less than or equal to 50%.

Applicant's summary and conclusion

Interpretation of results:

Category 2 (irritant)

Remarks:

Migrated information Criteria used for interpretation of results: EU

Executive summary:

The potential of Tinocat ES 96000 to cause dermal corrosion/irritation was assessed by a single topical application of 25 μL bulk volume (about 11 mg) of the test substance to a reconstructed three dimensional human epidermis model (EpiDerm™). For the corrosion test two EpiDerm™ tissue samples were incubated with the test substance for 3 minutes and 1 hour, respectively. The irritation test was performed with three EpiDerm™ tissue samples, 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 testsubstance treated epidermal tissues is compared to that of negative control tissues. The quotient of the values indicates the relative tissue viability. The EpiDerm™ skin corrosivity/irritation test showed the following results: The test substance is not able to reduce MTT directly. Corrosion test: The mean viability of the test-substance treated tissues determined after an exposure period of 3 minutes was 98%, and it was 92% after an exposure period of 1 hour. 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 32%. Based on the observed results and applying the evaluation criteria cited in chapter 3.8 it was concluded, that Tinocat ES 96000 shows a skin irritation potential in the EpiDerm™ skin corrosion/irritation test under the test conditions chosen.