Registration Dossier

Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2012-11-29 to 2012-11-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study reliable without restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)
Version / remarks:
adopted 2006-07-19
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 (incl. certificate)
Remarks:
signed 2009-03-30

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Vanadium, oxalate complexes
- Trivial name: vanadyl oxalate
- Physical state: blue crystals, odourless
- Storage condition of test material: at room temperature in tightly closed container (further detailed: Harlan CCR SOP SUBST.doc)

Test animals

Details on test animals and environmental conditions:
Not applicable - Since this is an in vitro study there is no information on test animals.

Test system

Vehicle:
unchanged (no vehicle)
Amount / concentration applied:
TEST MATERIAL
- Amount(s) applied (volume or weight with unit): approx. 500 mg of the test item were dispensed directly atop the bio-barrier.
Duration of treatment / exposure:
60 minutes
Observation period:
not applicable
Number of animals:
not applicable
Details on study design:
TEST SYSTEM
- Test kit:
The test kit (Name: Corrositex™; Supplier: Transia GmbH, 61239 Ober-Mörlen, Germany; Lot No.: CT052112) consists of a bio-barrier (collagen gel) and a chemical detection system separated by a cellulose membrane.
- Preparation of the bio-barrier:
One day prior to testing the bio-barrier matrix was prepared. The bio-barrier powder was solved in the bio-barrier diluent and heated for 20 ± 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 non-heated 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.

EXPERIMENTAL PERFORMEANCE
- Qualification Test: to test whether the test system is suitable for the test item, i.e. the test substance is detectable by the CDS, approx. 100 mg of the test item were transferred into the “Qualification Test Vial”. The vial was shaken until the solution appeared homogenous, and incubated for at least 1 minute. The colour change was recorded. Since a change in colour was visible in the “Qualification Test Vial”, the test item was considered to be suitable for the next step.
- Categorisation Test: qualified test substance is classified into categories (Category 1: strong acids or bases; Category 2: less strong acids and bases) by a screening test. The category, that a test substance is assigned to, will determine how the breakthrough time (if one occurs) will be interpreted. A test substance is assigned to a category based on its ability to induce a pH change in one of two defined buffers. One buffer is designed for detecting acids, the other for detecting bases.
The test item was categorised according to the pH value method to distinguish between weak and strong acids or bases as described in the Corrositex™ Instruction Manual (1995)*. The pH of the solution containing the test item in a “Category A Vial” and a “Category B Vial” was measured using pH sticks, where the Category A vial contained the acid buffer and Category B contained the base buffer. Test items with a solution pH that is ≤ 5 in the “Category A Vial” and ≥ 9 in the “Category B Vial”, i.e. strong acids or bases, were assigned to Category 1. Test items with a solution pH that is ≥ 5 in the “Category A Vial” and ≤ 9 in the “Category B Vial”, i.e. less strong acids and bases, were assigned to Category 2.
- Classification Test: 7 vials containing the CDS were acclimated at room temperature. A total of 4 vials was used for quadruplicate measurements of the test item, one vial was used as positive control (single measurement), and one vial was used as negative control (single measurement). One vial was used as colour reference for the CDS.
The prepared bio-barriers were placed on top of the CDS vials (not longer than 2 minutes prior to an application) and approx. 500 mg of the test item or 500 µL of the respective controls (positive control: sulfuric acid 95-97% (lot no. K40281631 929, Merck, 64295 Darmstadt, Germany); negative control: citric acid (lot no. 140986235008107, Sigma, 89555 Steinheim, Germany, 10% (w/v) solution in deionised water)) were applied per bio-barrier for 1 hour or 4 hours, depending on the results of the categorisation test. The time of the first colour change or precipitation in the CDS solution, i.e. bio-barrier penetration, was recorded.

INTERPRETATION OF THE RESULTS
Each vial was monitored for 60 minutes or 240 minutes depending on the results of the classification test, and the time of the first observable change in the CDS solution was recorded. The elapsed time between application and penetration of the bio-barrier was determined, and the mean of the quadruplicate measurements was calculated. The test item was categorised according to the criteria in table 1, which can be seen in the field "Any other information on materials and methods incl. tables" below.

ACCEPTABILITY OF THE ASSAY
The results were considered valid if the following acceptance criteria were met:
The test item induces a physical change (colour or precipitation) in the CDS solution in the qualification test, and the test substance is detectable by the CDS.
The negative control does not induce a change (i.e. colour or precipitation) in the CDS solution over 60 minutes in the classification test, and the functional integrity of the bio-barrier is demonstrated.
The positive control induces a change (i.e. colour or precipitation) in the CDS solution in the classification test after 0 – 3 minutes, and the breakthrough time of the positive control is within the acceptable range. The Corrositex™ assay will be accepted if the positive control time falls within ± two standard deviations of the positive control historical mean breakthrough time.

* Reference
- Corrositex™ instruction manual (1995). In Vitro International, 16632 Millikan Avenue, Irvine, CA 92606, USA.

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
other:
Value:
0
Remarks on result:
other:
Remarks:
Basis: other: minutes (mean of 4 replicates). Time point: after 60 minutes of treatment. Remarks: A change of colour of CDS reagent after treatment of bio-barriers with test item was not observed during the test, i.e. for 60 minutes. . (migrated information)

In vivo

Irritant / corrosive response data:
A change of colour of the CDS reagent after treatment of the bio-barriers with the test item was not observed for 60 minutes in any of the four replicates indicating that it did not break through the bio-barrier. The test item is not corrosive.

Any other information on results incl. tables

HISTORICAL DATA:

Table: Historical range of breakthrough times for the positive control

Historical mean time of first colour change of positive control [minutes]

1.31

Standard deviation [minutes]

0.45

Data of 46 studies performed at Harlan CCR from 2006 until 2012.

QUALIFICATION TEST:

The test item induced a change in colour in the qualification test after 1 minute of incubation. Since a change in colour was visible in the “Qualification Test Vial”, the test item was thus detected by the CDS and considered suitable for the next step.

CATEGORISATION TEST:

Since the pH value of the test item was 5 in the “Category A Vial” and 9 in the “Category B Vial”, the test item does not appear to be a strong acid or base and was assigned to category 2. It was concluded that the required duration of the classification test was 60 minutes.

CLASSIFICATION TEST:

The negative control did not penetrate the bio-barrier, and thus the bio-barrier was considered functionally integer.

The positive control, i.e. a colour change, was detected by the CDS after 1 minute, and thus the breakthrough time of the positive control fell within ± two standard deviations of the positive control historical mean breakthrough time.

A change in the CDS was not observed for 60 minutes in any of the four replicates of the test item indicating that it did not break through the bio-barrier.


Test Group

Time Interval of
Colour Change

UN Packaging Group

R-Sentence

GHSand Regulation (EC) No 1272/2008 (CLP)

Negative Control

Colour change was not observed for 60 minutes

-

-

-

Positive Control

1 minute

I

R35

1A

Test Item

Colour change was not observed for 60 minutes

Non-corrosive

-

-

All validity criteria of the test were met. No other effects were observed in any of the treatments.

 

Applicant's summary and conclusion

Interpretation of results:
other: the test item is not corrosive.
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Vanadium, oxalate complexes is not corrosive to skin according to the EC-Commission directive 67/548/EEC and EC-Regulation 1272/2008 and subsequent regulations.