Registration Dossier

Diss Factsheets

Toxicological information

Skin irritation / corrosion

Currently viewing:

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
6th February 2019 - 8th February 2019
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2019
Report date:
2019

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)
Version / remarks:
Adopted 29th July 2016
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EC Guideline No. 440/2008. Part B: Methods for the Determination of Toxicity and other health effects, Guideline B.40 BIS: "In Vitro Skin Corrosion: Human Skin Model Test".
Version / remarks:
Official Journal of the European Union No. L142, 31 May 2008.
Deviations:
no
GLP compliance:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
2,9,16,23-tetrakis(2,5-di-tert-butyl-4-methoxyphenoxy)phthalocyanine
EC Number:
831-167-5
Cas Number:
2126827-44-7
Molecular formula:
C92H106N8O8
IUPAC Name:
2,9,16,23-tetrakis(2,5-di-tert-butyl-4-methoxyphenoxy)phthalocyanine
Test material form:
solid
Specific details on test material used for the study:
Batch (Lot) Number: 2715/18/01
Expiry date: 01 July 2020
Physical Description: Dark green solid
Purity/Composition: 99%
Storage Conditions: At room temperature protected from light

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Cell source:
other: Cultured to form a multilayered, highly differentiated model of the human epidermis
Source strain:
other: Not applicable
Details on animal used as source of test system:
EpiDerm Skin Model (EPI-200, Lot no.: 29944 Kit I and J).
Justification for test system used:
Recommended system according to the OECD 431 and EC TG.
Vehicle:
unchanged (no vehicle)
Details on test system:
The model consists of normal, human-derived epidermal keratinocytes which have been cultured to form a multilayered, highly differentiated model of the human epidermis. It consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum containing intercellular lamellar lipid layers arranged in patterns analogous to those found in vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of 10 mm cell culture inserts.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
yes, concurrent MTT non-specific colour control
Amount/concentration applied:
Substance: 25.4 to 41.6 mg per tissue
Positive control: 50 μL per tissue
Negative control: 50 μL per tissue
Duration of treatment / exposure:
3 minutes and 60 minutes
Duration of post-treatment incubation (if applicable):
3 hours at 37°C in air containing 5% CO2.
Number of replicates:
Duplicate

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
% tissue viability
Remarks:
3-minute time point
Run / experiment:
mean of two samples
Value:
ca. 94
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: No indication of corrosion
Irritation / corrosion parameter:
% tissue viability
Remarks:
60 minute time point
Run / experiment:
Mean of two samples
Value:
ca. 100
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: No indication of corrosion
Other effects / acceptance of results:
The non-specific reduction of MTT by the substance was -7.4% and 2.2% of the negative control tissues after 3 minutes and 1 hour, respectively.
In addition to the normal 3-minute and 1-hour procedure, two tissues were treated with substance instead of MTT solution these tissues were incubated with DMEM. The color interference by the substance was 5.4% and 7.3% of the negative control tissues after 3 minutes and 1 hour respectively.
For the true tissue viability to be calculated, in addition to the normal 3-minute and 1-hour procedure, two freeze-killed tissues treated with test item were used but instead of MTT solution these tissues were incubated with DMEM. The nonspecific color in freeze-killed tissues by the substance was 2.8% and 4.4% of the negative control tissues after 3 minutes and 1 hour respectively.

Any other information on results incl. tables

Mean Absorption in the in vitro Skin Corrosion Test

 

 3 -minute application viability (%)

   1 -hour application viability (%)

A (OD570)

B (OD570)

Mean (OD570)

SD (+/-)

A (OD570)

B (OD570)

Mean (OD570)

SD (+/-)

Negative control

1.375

1.511

1.443

0.097

1.316

1.538

1.427

0.157

Substance*

1.279

1.423

1.351

0.101

1.281

1.562

1.422

0.199

Positive control

0.511

0.324

0.087

0.132

0.250

 0.117

0.183

0.094

SD = Standard deviation

Duplicate exposures are indicated by A and B.

* For the 3 minute exposure the substance values are corrected for the color interference (5.4%).

For the 1 hour exposure the substance values are corrected for the non-specific MTT reaction and color interference (2.2%

and 7.3 %, respectively). In addition these values are corrected for the nonspecific color in freeze-killed tissues (4.4%).

The values are corrected for background absorption (0.0475). Isopropanol was used to measure the background absorption.

Mean Tissue Viability in the in vitro Skin Corrosion Test

 

 

3 -minute application viability (% of control)

1 -hour application viability (% of control)

 Negative control 100 100 
 Substance 94  100 
 Positive control 29  13

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The substance is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this study.
Executive summary:

In a reliable in vitro skin corrosion study, conducted according to the OECD Guideline 431, 'In Vitro Skin Corrosion: reconstructed human epidermis (RHE) test method', the undiluted substance ( 25.4 to 41.6 mg) was applied onto reconstructed human skin tissue (epidermal model, EpiDerm tissue (0.6 cm²)) in duplicate for a period of 3 or 60 minutes.

 

Skin corrosion is expressed as the remaining cell viability after exposure to the substance. The relative mean tissue viability obtained after 3-minute and 1-hour treatments with the substance compared to the negative control tissues was 94 % and 100 %, respectively. Because the mean relative tissue viability for the substance was not below 50% after the 3-minute treatment and not below 15% after the 1-hour treatment the substance is considered to be non corrosive.

 

In conclusion, the substance is not corrosive in the in vitro skin corrosion test under the experimental conditions described in this study.