Tetraol

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

2014-11-25 to 2015-05-20

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

26 July 2013

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Test material form:

solid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Lot/batch No.of test material: 003
- Expiration date of the lot/batch: 01 Jan 2020

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, exclusion of direct sunlight, test substance is hygroscopic
- Stability under test conditions: stable

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Vehicle:

water

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™
- Tissue batch number: 21604

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: room temperature

REMOVAL OF TEST MATERIAL AND CONTROLS
- Number of washing steps: once after treatment and once after MTT incubation
- Observable damage in the tissue due to washing: no
- Modifications to validated SOP: no

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1 mg/mL
- Incubation time: 3 hours
- Spectrophotometer: SunriseTM Absorbance Reader
- Wavelength: 570 nm
- Filter: without reference filter

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability: 1.821±0.108 (Acceptance criteria: OD (540-570 nm) [1.0 - 3.0])
- Barrier function: 5.32 hrs (Acceptance criteria: ET-SO [4.77 - 8.72 hrs])
- Contamination: sterile (acceptance criteria: no contamination)

NUMBER OF REPLICATE TISSUES: 1

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- killed tissues
- Procedure used to prepare the killed tissues: tissues were freeze killed
- No. of replicates: 2
- Method of calculation used: 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.

NUMBER OF INDEPENDENT TEST SEQUENCES / EXPERIMENTS TO DERIVE FINAL PREDICTION: 2

PREDICTION MODEL / DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50 %, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15 %
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is greater than or equal to 15 %.

Control samples:

yes, concurrent vehicle

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

TEST MATERIAL
- Amount applied: 25 µL

VEHICLE
- Amount applied: 25 µL

NEGATIVE CONTROL
- Amount applied: 50 µL de-ionized water

POSITIVE CONTROL
- Amount applied: 50 µL
- Concentration: 8 N potassium hydroxide

Duration of treatment / exposure:

3 min and 1 hour

Number of replicates:

2

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min exposure

Value:

102.4

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 hours exposure

Value:

107.6

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: No
- Direct-MTT reduction: Test substance is not able to reduce MTT directly.
- Colour interference with MTT: Not applicable

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: Yes
- Acceptance criteria met for positive control: Yes
- Acceptance criteria met for variability between replicate measurements: Yes

Any other information on results incl. tables

Table 1: 3 min exposure

Test substance		Tissue 1	Tissue 2	Mean	SD	CV [%]
NC	Mean OD570	2.007	1.967	1.987
	Viability [% of NC]	101.0	99.0	100.0	1.4	1.4
Test substance	Mean OD570	1.963	2.107	2.035
	Viability [% of NC]	98.8	106.1	102.4	5.1	5.0
PC	Mean OD570	0.378	-	0.378
	Viability [% of NC]	19.0	-	19.0

Table 2: 1 hour exposure

Test substance		Tissue 1	Tissue 2	Mean	SD	CV [%]
NC	Mean OD570	2.031	2.032	2.031
	Viability [% of NC]	100.0	100.0	100.0	0.02	0.02
Test substance	Mean OD570	2.093	2.279	2.186
	Viability [% of NC]	103.0	112.2	107.6	6.5	6.0
PC	Mean OD570	0.113	0.089	0.101
	Viability [% of NC]	5.5	4.4	5.0	0.8	16.5

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met