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Skin irritation / corrosion

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Administrative data

Endpoint:
skin irritation / corrosion
Remarks:
in vitro
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: well performed OECD study with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)
Deviations:
no
Qualifier:
according to
Guideline:
other: ICCVAM Minimum Performance Standards: In Vitro Membrane Barrier Test Systems for Skin Corrosion, June 23, 2003
Deviations:
no
Qualifier:
according to
Guideline:
other: ICCVAM Recommended Performance Standards for in vitro Test Methods for Skin Corrosion (May 2004)
Deviations:
no
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
purity; 89.7 % (w/w) (main component)

Test system

Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): Approximately 500 mg of the neat test item were dispensed directly atop the bio-barrier.


Duration of treatment / exposure:
The prepared bio-barriers were placed atop the CDS vials (not longer than 2 min prior to application) and approximately 500 mg of the test item or each 500 µL of the controls were applied per bio-barrier for 1 hour or 4 hours, depending on the results of the categorisation test.
Observation period:
Qualify Test
In order to test whether the test system is suitable for the test item, approximately 100 mg of the test item were applied into the “Qualify Test Vial”. The vial was shaken until the solution appeared homogenous, and incubated for 1 minute. Afterwards, the colour change was noted.

Categorisation Test

In the categorisation test the observation period of the test item after application to the bio-barrier was determined:
• Category 1: observation period after application will be 4 hours
• Category 2: observation period after application will be 1 hour





 
Details on study design:
Test Kit:
Name: Corrositex™
Supplier: Transia GmbH, 61239 Ober-Mörlen, Germany
Lot No.: CT051214

Preparation of the Bio-barrier:
One day of testing the bio-barrier matrix was prepared. The bio-barrier powder was solved in the bio-barrier diluent and heated for 20 plus/minus 2 minutes at 68 – 70 °C in a water bath under continuous stirring. The temperature did not exceed 70 °C. The mixture was allowed to cool in the turned-off water bath for another 10 minutes. The mixture was then filled into the membrane holders (200 µL per membrane holder). Air bubbles were avoided. The filled membrane holders were sealed with parafilm and were stored at 2 – 8 °C until further use.

Preparation of the Test Item:
Approximately 500 mg of the neat test item were dispensed directly atop the bio-barrier.

Negative Control
Citric acid (10% (w/v) solution in deionised water) was used as negative control. 500 µL were applied to only one bio-barrier.

Positive Control
Sulfuric acid 95-97% was used as positive control. 500 µL were applied to only one bio-barrier.

Experimental Performance

Qualify Test:
In order to test whether the test system is suitable for the test item, approximately 100 mg of the test item were applied into the “Qualify Test Vial”. The vial was shaken until the solution appeared homogenous, and incubated for 1 minute. Afterwards, the colour change was noted.

Categorisation Test:
In the categorisation test the observation period of the test item after application to the bio-barrier was determined:
Category 1: observation period after application will be 4 hours
Category 2: observation period after application will be 1 hour

Since the test item is strongly coloured the categorisation of the test item has to be done as follows. A 10% solution has to be prepared in deionised water. The pH value of the solution has to be determined. Each 150 µL of the solution has to be added to the “Category A Vial” in case of an pH value < 7 or to the “Category B Vial” in case of a pH value > 7. The vial has to be closed, shaken, and the pH value has to be determined again. Test items showing a pH value ≤ 5 in the “Category A Vial” have to be assigned to category 1 and test items showing a pH value of ≥ 5 have to be assigned to category 2. Test items showing a pH value of ≥ 9 in “Category B Vial” have to be assigned to category 1 and test items showing a pH value of ≤ 9 have to be assigned to category 2.

Classification Test:
7 vials containing the CDS were pre-warmed to room temperature. 4 vials were used for quadruplicate measurement of the test item, the vial labelled (+) was used for the positive control (single measurement), and the vial labelled (-) was used for the negative control (single measurement). The vial labelled C was used as colour reference for the CDS.

Application of the test item formulations and the controls was performed staggered to ensure accurate reaction times to be recorded.
The prepared bio-barriers were placed atop the CDS vials (not longer than 2 min prior to application) and approximately 500 mg of the test item or each 500 µL of the controls were applied per bio-barrier for 1 hour or 4 hours, depending on the results of the categorisation test. The time interval of the possible colour change or precipitation in the CDS solution was recorded in the raw data file.

Data Recording:
The data generated were recorded in the laboratory protocol. The results were presented in tabular form, including experimental groups with the test item, negative, and positive controls.


Results and discussion

In vivo

Irritant / corrosive response data:
Not corrosive to skin.

Any other information on results incl. tables

Qualify Test

The test item induced a change in colour in the qualify test after 1 minute incubation. Since a change in colour was visible in the “Qualify Test Vial”, the test item was considered to be suitable for the next step.

Categorisation Test

Since the test item was deeply coloured (dark purple) the categorisation test was done by pH value determination. The test item solution showed pH value of > 7 and was therefore added in thecategory B vial. Since the pH of the solution in the category B vial reached a value of ≤ 9, the test item was assigned to category 2. Additionally, the test item solution was added in the category A vial, since the pH value was ≥ 5, the assigning of the test item to category 2 was confirmed.

Classification Test

 


Test Group

Time Interval of
Colour Change

UN Packaging Group

R-Sentence

GHS

Negative Control

Colour change was not observed after > 60 minutes

Non corrosive

-

-

Positive Control

1 minute 40 seconds

I

R35

1A

Test Item

> 60 minutes

Non corrosive

-

-

The test item was classified as non corrosive. 

Applicant's summary and conclusion

Interpretation of results:
other: does not meet criteria for classificaion as corrosive according to EU / does not meet criteria for classificaion as corrosive according to UN GHS / non-corrosive according to UN Packaging Group (DOT)
Remarks:
Criteria used for interpretation of results: other: EU / UN GHS / UN Packaging Group (DOT)
Conclusions:
In conclusion, it can be stated that in this study and under the experimental conditions reported, the test item

- does not meet criteria for classificaion as corrosive according to EU,
- does not meet criteria for classificaion as corrosive according to UN GHS,
- is non-corrosive according to UN Packaging Group (DOT).
Executive summary:

RESULTS AND DISCUSSION

Qualify Test

The test item induced a change in colour in the qualify test after 1 minute incubation. Since a change in colour was visible in the “Qualify Test Vial”, the test item was considered to be suitable for the next step.

Categorisation Test

Since the test item was deeply coloured (dark purple) the categorisation test was done by pH value determination. The test item solution showed pH value of > 7 and was therefore added in thecategory B vial. Since the pH of the solution in the category B vial reached a value of ≤ 9, the test item was assigned to category 2. Additionally, the test item solution was added in the category A vial, since the pH value was ≥ 5, the assigning of the test item to category 2 was confirmed.

Classification Test

Test Group

Time Interval of Colour Change

UN Packaging Group

R-Phrase

GHS

Negative Control

Colour change was not observed after > 60 minutes

Non corrosive

-/-

-/-

Positive Control

1 minute 40 seconds

I

R35

1A

Test Item

> 60 minutes

Non corrosive

-/-

-/-

The test item was classified as non corrosive. 

Discussion

This in vitrostudy was performed to assess the corrosive potential of the test item by means of theIn VitroMembrane Barrier Test for Skin Corrosion using the Corrositex™ test kit.

The test item passed the qualify test and was considered as suitable for testing with the Corrositex™ test kit.

Since the test item was deeply coloured the categorisation test was done be pH value determination. The test item showed a pH value of ≤ 9 in theCategory B vial and was assigned to category 2.

Approx. 500 mg of the test item were applied on quadruplicate bio-barriers for the classification test. The bio-barriers were placed atop vials containing the CDS reagent. The time needed to corrode the bio-barrier and induce a distinct change in the colour of the CDS reagent was monitored for each of the vials.

500 µL of the negative control (10% citric acid) was placed atop one bio-barrier and the colour change of the CDS reagent was monitored during a period of > 60 minutes.

500 µL of the positive control (Sulfuric acid 95-97%) was placed atop one bio-barrier and the colour change of the CDS reagent was monitored.

The negative control did not induce a change in the colour of the CDS reagent after an observation period of > 60 minutes. The positive control showed a distinct change in the colour of the CDS reagent in the time interval of 1 minute 40 seconds. These results ensured the validity of the test.

There was no change of colour of the CDS reagent after treatment of the bio-barriers with the test item during the observation period of > 60 minutes. According to the classification criteria given the test item was classified as not corrosive.