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Toxicological information

Skin irritation / corrosion

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Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 25 JUNE 2018 to 31 AUGUST 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report date:
2018

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Deviations:
no
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
1-butyl-5-[(4-chlorophenyl)azo]-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile
EC Number:
274-126-1
EC Name:
1-butyl-5-[(4-chlorophenyl)azo]-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile
Cas Number:
69808-32-8
Molecular formula:
C17H17ClN4O2
IUPAC Name:
1-butyl-5-[(4-chlorophenyl)azo]-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile
Test material form:
solid: particulate/powder
Details on test material:
- State of aggregation: powder
- Particle size distribution: not determined
- Mass median aerodynamic diameter (MMAD): not determined
- Geometric standard deviation (GSD): not determined
- Shape of particles: not determined
- Surface area of particles: not determined
- Crystal structure: not determined
- Coating: none
- Surface properties: not determined
- Density: 1.3526 ± 0.0010 g/cm3 at 20.0 ± 0.2°C
- Moisture content: <0.5%
- Residual solvent: not detected
- Activation: none
- Stabilisation: none
- Other:

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
foreskin from multiple donors
Source strain:
not specified
Details on animal used as source of test system:
No animals were used
Justification for test system used:
The objective of this study was to determine the potential skin corrosivity of a test item 1-butyl-5-[(4-chlorophenyl)azo]-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile with the in vitro skin corrosion test using reconstructed human epidermis (RhE; EpiSkinTM-SM) following OECD test guideline 431. This method allows classification of the test item in non-corrosive or corrosive substances and mixtures according to the United Nations (UN) Globally Harmonized System of Classification and Labelling of Chemicals (GHS). It also allows a partial sub-categorization of corrosives.
Vehicle:
unchanged (no vehicle)
Details on test system:
RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: Reconstructed Human Epidermis (RhE; EpiSkinTM-SM).
- Tissue batch number(s): 18-EKIN-030
- Production date:
- Shipping date:
- Delivery date:
- Date of initiation of testing: 24.07.18

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 +/- 1ºC
- Temperature of post-treatment incubation (if applicable):

REMOVAL OF TEST MATERIAL AND CONTROLS
-Volume and number of washing steps:
- Observable damage in the tissue due to washing:
- Modifications to validated SOP:

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 0.3 mg/ml
- Incubation time: 3h +/- 15 min
- Spectrophotometer:
- Wavelength: between 540 to 600 nm
- Filter: no reference filter was used
- Filter bandwidth:
- Linear OD range of spectrophotometer:

FUNCTIONAL MODEL CONDITIONS WITH REFERENCE TO HISTORICAL DATA
- Viability:
- Barrier function:
- Morphology:
- Contamination:
- Reproducibility:

NUMBER OF REPLICATE TISSUES:

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues
- Procedure used to prepare the killed tissues (if applicable):
- N. of replicates :
- Method of calculation used:

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

PREDICTION MODEL / DECISION CRITERIA (choose relevant statement)
- The test substance is considered to be corrosive to skin if [complete, e.g. 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 [complete, e.g. 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%.]
- Justification for the selection of the cut-off point(s) if different than recommended in TG 431 and 439:
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 20g
- Concentration (if solution):

VEHICLE
- Amount(s) applied (volume or weight with unit): 100%
- Concentration (if solution):
- Lot/batch no. (if required):
- Purity: none

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL of negative control (0.9% NaCl)
- Concentration (if solution): 0.9% NaCl

POSITIVE CONTROL
- Amount(s) applied (volume or weight):50 µL of positive control (glacial acetic acid)
- Concentration (if solution): 100%
Duration of treatment / exposure:
3 min, 1h (±5 min) and 4h (±10 min) at room temperature
Duration of post-treatment incubation (if applicable):
incubated for 3h (± 15 min) under culture conditions
Number of replicates:
A single testing run was sufficient becuuse was unequivocal.

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
Run 1 (duration exposure time 3 minutes), Run 2 (duration exposure time 1 hour) and run 3 (duration exposure time 4 hours)
Value:
>= 35
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
no indication of irritation
Remarks:
Taking into account the results obtained in the current study, it is concluded that, under the assayed experimental conditions and according to the OECD guideline for the Testing of Chemicals Nº. 431, the test item 1-butyl-5-[(4-chlorophenyl)azo]-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile can be categorised as Non-Corrosive according to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Taking into account the results obtained in the current study, it is concluded that, under the assayed experimental conditions and according to the OECD guideline for the Testing of Chemicals Nº. 431, the test item 1-butyl-5-[(4-chlorophenyl)azo]-1,2-dihydro-6-hydroxy-4-methyl-2-oxonicotinonitrile can be categorised as Non-Corrosive according to the United Nations Globally Harmonized System of Classification and Labelling of Chemicals.