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Diss Factsheets

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: UN GHS (2003, last rev. 2015)
Principles of method if other than guideline:
Based on a “Statement on the Scientific Validity of In Vitro Tests for Skin Irritation” of the European Commission (November 2008), official acceptance of the test method in the EU was achieved and implemented in EU, 2008a, Council Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to EC Regulation No 1907/2006 of the European Parliament and of the Council on REACH; 1st ATP 2009: EC Regulation No 761/2009 of 23 July 2009 amending, for the purpose of its ATP, EC Regulation No 440/2008 laying down test methods pursuant to EC Regulation No 1907/2006 of the European Parliament and of the Council on REACH, section B46.
GLP compliance:
yes (incl. QA statement)
Remarks:
Date of inspection: 13-16 July 2015, Date of Signature: 14 September 2015

Test material

Constituent 1
Chemical structure
Reference substance name:
1-(methylamino)anthraquinone
EC Number:
201-417-2
EC Name:
1-(methylamino)anthraquinone
Cas Number:
82-38-2
Molecular formula:
C15H11NO2
IUPAC Name:
1-(methylamino)anthraquinone
Test material form:
solid

In vitro test system

Test system:
human skin model
Vehicle:
unchanged (no vehicle)
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
Each approximately 25 mg (~ 39 mg/cm2 according to guideline) of the test item were applied to the tissues, wetted with 25 µL of DPBS, and spread to match the surface of the tissue for a complete treatment time of 60 minutes.
Duration of treatment / exposure:
60 minutes.

Test animals

Species:
other: reconstituted human epidermis model

Test system

Type of coverage:
other: Topical
Preparation of test site:
other: Not applicable
Vehicle:
other: No vehicle used
Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit):
Each approximately 25 mg (~ 39 mg/cm2 according to guideline) of the test item were applied to the tissues, wetted with 25 µL of DPBS, and spread to match the surface of the tissue

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL DPBS (MaTek) on each tissue


POSITIVE CONTROL
- Amount(s) applied (volume or weight): 30 µL of 5% SLS on each tissue
Duration of treatment / exposure:
60 minutes
Observation period:
Not applicable
Number of animals:
Not applicable

Results and discussion

In vitro

Resultsopen allclose all
Irritation / corrosion parameter:
other: Mean relative absorbance in %
Remarks:
MTT-Test
Run / experiment:
all
Value:
ca. 104.4
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
all
Value:
ca. 97.2
Vehicle controls validity:
valid
Negative controls validity:
valid
Positive controls validity:
valid

In vivo

Irritant / corrosive response data:
Compared to the relative absorbance value of the negative control the mean relative absorbance value was reduced to 8.8% after exposure of the skin tissues to the test item. This value is below the threshold for irritancy of ≤ 50%. Therefore, the test item is considered to possess an irritant potential.
Other effects:
No

Any other information on results incl. tables

Results after treatment with Solvaperm-Rot PFS and the controls (exposure interval of 60 minutes):

 Dose Group  Tissue No.:

Absorbance

570 nm

Well 1

 Absorbance

570 nm

Well 2

 Absorbance

570 nm

Well 3

 Mean

Absorbance

of 3 Wells

 Mean Absorbance

of 3 wells

blank corrected

 Mean Absorbance

of 3 wells

after blank correction

Relative

Absorbance [%]

Tissue 1,2,3 *

Relative Standard

Deviation [%] 

Mean Relative Absorbance 

[% of Negative Control]**

Step 2*** Mean Rel. Absorbance [% of Negative Control] after colour interference and MTT reduction correction 
 Blank   0.036  0.037   0.036  0.036  0.000            
 Negative Control  1  1.770 1.757  1.747  1.758   1.721

 

1.710      

100.6         
   2  1.852  1.733  1.745 1.777   1.739 101.7  2.1  100.0     
   3 1.748   1.700 1.675  1.708  1.670  97.7         
 Positive Control  1  0.119 0.116  0.116  0.117   0.079

  1.072      

 4.6        
   2 0.107  0.111   0.107 0.108  0.071  4.1  9.4  4.2     
   3 0.103  0.104  0.104   0.104 0.066  3.9         
 Test item  1 1.846  1.812   1.809  1.822 1.785 

 

1.784      

 104.4        
   2 1.921   1.902 1.915  1.912   1.875 109.6   5.1 104.4  1.663  97.2 
   3  1.755  1.728  1.711 1.731  1.694  99.1         
 Blank   0.037  0.038  0.038  0.038  0.000             

 Negativ Control

without MTT

0.041  0.046  0.040  0.042  0.005         100.0    

 Test item

without MTT

 1 0.166   0.154 0.155  0.158  0.121         2666.2    

*        relative absorbance per tissue [rounded values]

**      relative absorbance per treatment group [rounded values]

*** Mean ODtest item minus ODtest item without MTT

The optical pre-experiment (colour interference pre-experiment) to investigate the test item’s colour change potential in water led to a change in colour. An additional test with one viable tissue was necessary. The result was used for data correction of the results in the main experiment.

Optical evaluation of the MTT-reducing capacity of the test item after 1 hour incubation with MTT-reagent did not show blue colour. An additional test with freeze-killed tissues was not necessary.

The corrected mean relative absorbance value of the test item after correction, corresponding to the cell viability, were not reduced (97.2%; threshold for irritancy:50%) compared to the result of the negative control, consequently the test item was not irritant to skin. 

Applicant's summary and conclusion

Interpretation of results:
other: not irritant
Remarks:
Migrated information Criteria used for interpretation of results: OECD GHS
Conclusions:
In conclusion, it can be stated that in this study and under the experimental conditions reported, Solvaperm-Rot PFS is not irritant to skin according to UN GHS and EU CLP regulation.
Executive summary:

This in vitro study was performed to assess the irritation potential of Solvaperm-Rot PFS by means of the Human Skin Model Test.

The test item passed the MTT interference pre-test. Due to its intensive colour, an additional test with one viable tissue (without MTT addition) was necessary to correct the result in the main experiment.

Approximately 25 mg of the test item were applied to each tissue, wetted with 25 µL of DPBS, and spread to match the surface of triplicate tissue.

30 µL of either the negative control (DPBS) or the positive control (5% SLS) were applied to triplicate tissue each.

The test item and the positive and negative controls were washed off the skin tissues after 60 minutes treatment. After further incubation for about 42 hours the tissues were treated with the MTT solution for 3 hours following approximately 69 hours extraction of the colorant from the cells. The amount of extracted colorant was determined photometrically at 570 nm.

After treatment with the negative control the absorbance values were well within the required acceptability criterion of mean OD³0.8 and ≤ 2.8 for the 60 minutes treatment interval thus showing the quality of the tissues.

Treatment with the positive control induced a decrease in the relative absorbance as compared to the negative control to 4.2% thus ensuring the validity of the test system.

The relative standard deviations between the % variability values of the test item, the positive and negative controls in the main test were below 10% (threshold of the "OECD Guideline for the Testing of Chemicals 439:In vitroSkin Irritation: Reconstructed Human Epidermis Test Method”: < 18%), thus ensuring the validity of the study.

Compared to the relative absorbance value of the negative control the corrected mean relative absorbance value was slightly reduced (97.2%) after exposure of the skin tissues to the test item. This value is above the threshold for irritancy of ≤ 50%. Therefore, the test item is not considered to possess an irritant potential.

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