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Administrative data

Endpoint:
skin corrosion: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
27 Feb 2019 - 21 May 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 guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)
Version / remarks:
28 July 2015
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
2-[2-(dimethylamino)ethoxy]-N-methylethanamine
EC Number:
641-132-4
Cas Number:
93240-93-8
Molecular formula:
C7 H18 N2 O
IUPAC Name:
2-[2-(dimethylamino)ethoxy]-N-methylethanamine
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Name of test substance: Ethanamine,N,N-dimethyl-2-[2-(methylamino)ethoxy]-
- Test substance No.: 19/0034-1
- Source and lot/batch No.of test material: 08324216K0
- Expiration date of the lot/batch: 01 Nov 2020
- Purity: 86.6 corrected area-%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, under N2
- Solubility and stability of the test substance in the solvent/vehicle: The stability under storage conditions over the study period was
guaranteed by the sponsor, and the sponsor holds this responsibility.

OTHER SPECIFICS:
pH value: 12.3 (10% (mass), information given in the material safety data sheet) :
Physical state / color: Liquid / colorless, clear

In vitro test system

Test system:
artificial membrane barrier model
Details on test system:
The Corrositex® assay is a standardized in vitro corrosion test. The Corrositex® assay kit is
available commercially from InVitro International.
The Corrositex® bio barrier membrane is a test system consisting of a reconstituted collagen matrix.
The assay is based on the time that the test substance requires to penetrate through
the Corrositex® bio barrier membrane and produce a change in the Chemical Detection System (CDS).
The Corrositex® assay is used to determine the corrosive potential of test substances. The
assay is limited to testing materials that cause detectable pH changes in the CDS.

Experimental procedure
The experimental design of this study consisted of a qualification screen with the CDS(to determine if
a color change can be detected) and a categorization screen (to categorize weak acids/bases and
strong acids/bases), which were performed as a pretest (experimentalconduct in accordance with
GLP, but without a GLP status) and a definitive Corrositex® assay.

The Corrositex® assay was evaluated on the basis of the color change of the CDS. The time until a
color change was observed was recorded manually, and the breakthrough times of the four replicates
were used to determine the corrosive potential of the test substance.

Test substance compatibility with the assay (qualification screen)
For the qualification screen, 150 μL of the test substance were added to the CDS screening tube. If
the test substance failed to produce a color change in the CDS within one minute, the test substance
could not be analyzed in this system, and no further testing was required.

Categorization screen
The categorization screen was used to assess the appropriate scoring scale for the test substance.
The categorization screen was performed by adding 150 μL of test substance to tube A and B each.
Each tube was mixed, and the resulting color was observed. If required, 2 drops of the "confirm"
reagent were added to tube B, the tube was mixed, and the resulting color was observed. The
categorization kit and color chart provided by InVitro International were used to determine the
category.
The test substance was scored as category 1 (high acid/alkaline reserve) or category 2 (low
acid/alkaline reserve)

Bio barrier preparation
The vial containing the bio barrier matrix powder was placed in a water bath at 64 – 68ºC. The entire
content of the bio barrier diluent vial was added slowly to the matrix powder. The stir bar rotated
slowly enough to avoid foaming of the solution. 200 μL solubilized matrix were pipetted into each of
the membrane discs. The membrane discs were then refrigerated for at least 2 hours at 2 – 8°C.
The bio barriers were wrapped and stored at 2 – 8°C for a maximum of 7 days.
Any remaining matrix solution was stored at 2 – 8°C for up to 30 days in order to prepare
additional bio barrier membrane discs.

Corrositex® assay
Following the acceptance of the positive control, the Corrositex® assay was performed for the
test substance. Four vials containing the CDS were used for the test substance.
In addition, one vial was used for the PC, the NC and the color (blank) control each. A membrane disc
coated with the bio barrier matrix was placed into one vial containing the
CDS. 500 μL undiluted test substance were added onto the membrane disc. An electronic time clock
was started with the application. The vial was observed for three minutes for any change in the CDS.

If no color change was observed within three minutes, the remaining membranes were treated
with the test substance. An electronic time clock was started with each application. The vials were
observed continuously for the first ten minutes. Thereafter, the vials were observed for approximately
ten minutes around the time points relevant for evaluation or until breakthrough of the test substance
occurred. The elapsed time between test substance application and the first change in the indicator
solution (i.e. barrier penetration) was recorded.

The positive control vial was prepared as described above and contained one pellet of sodium
hydroxide on top of the membrane disc. This vial was monitored continuously until
breakthrough occurred.

The negative control vial was prepared as described above and contained 500 μL 10% citric acid.
This vial was observed for 60 minutes and evaluated as “non-corrosive” if no reaction could be observed.

Acceptance criteria
The Corrositex® assay was accepted if the breakthrough time for the positive control substance was
in the historic control range (mean ± 2-3x standard deviations, In order to demonstrate the functional
integrity of the membrane barrier, the acceptance criterium for the negative control was to not induce
membrance breakthrough within a 60 min observation period.
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
500 μL undiluted test substance
Duration of treatment / exposure:
60 minutes
Number of replicates:
6

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
penetration time (in minutes)
Run / experiment:
mean
Value:
45.16
Vehicle controls validity:
not applicable
Negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: The mean breakthrough time determined in the in vitro membrane barrier test, considering 6 valid vials of two test runs

Any other information on results incl. tables

Two test runs of the Corrositex®Skin Corrosion Test were performed.

In the first test run high variability between the single vials were noted (single break through times see Table 1).

In order to clarify the result of the first test run, a second test run was performed.

In the second test run all four vials showed homogeneous break through times in the range of vials 3 and 4 of the first test run

(single break through times see Table 2).

Thus, the results of vial 1 and 2 of the 1sttest run are considered not to be valid and are excluded from evaluation.

 The times recorded per test run are shown in the following tables:

 

Table 1: 1sttest run; breakthrough times of the test substance and the PC and NC

 

 

Test substance

Break Through Time [min:s]

Vial 1 *

Vial 2 *

Vial 3

Vial 4

Mean **

19/0034-1

2:36

19:10

43:42

43:47

43:45

Controls:

 

PC:

Sodium hydroxide, solid

16:06

-

-

-

-

NC:

10% citric acid

NB

-

-

-

-

* outlier, excluded from evaluation

** mean of vial 3 and 4

NB = no breakthrough within maximum observation period (60 min)

 

Table 2: 2ndtest run; breakthrough times of the test substance and the PC and NC

 

 

Test substance

Break Through Time [min:s]

Vial 1

Vial 2

Vial 3

Vial 4

Mean

19/0034-1

44:05

47:27

47:33

48:04

46:47

Controls:

 

PC:

Sodium hydroxide, solid

15:30

-

-

-

-

NC:

10% citric acid

NB

-

-

-

-

 

NB = no breakthrough within maximum observation period (60 min)

Applicant's summary and conclusion

Interpretation of results:
Category 1C (corrosive) based on GHS criteria
Conclusions:
Based on the results observed and by applying the evaluation criteria described in chapter 3.8., it was concluded that
Ethanamine, N,N-dimethyl-2-[2-(methylamino)ethoxy]- shows a corrosive potential in the Corrositex® Skin Corrosion Test
under the test conditions chosen.The mean breakthrough time determined in the in vitro membrane
barrier test, considering 6 valid vials of two test runs, was 45 minutes and 16 seconds.
The breakthrough time indicates that the test substance has a weak corrosive potential and should be assigned to UN GHS skin corrosivity subcategories 1C or UN Transport Packing Group III as specified in OECD TG 435 and Instruction Manual.
Executive summary:

The Corrositex®assay showed the following results:

The qualification screen demonstrated that the test substance is able to react with the CDS and produce a visible color change.

Therefore, the membrane barrier test method was determined to be suitable for the evaluation of the corrosive potential

of the test substance.

A timescale category test was carried out to distinguish between weak and strong acids or bases. The test substance was assigned to timescale category 2.

In each test run of the main test four Corrositex®bio barrier membranes were treated with the undiluted test substance.

In the first test run high variability between the single vials were noted (single break through times [min:sec]: 2:36, 19:10, 43:42 and 43.47).

Two vials out of the four vials treated with the test substance yielded considerably shorter break through times than the other two vials.

Especially the first vial showed an extremly shorter break through time. Thus, a technical fault was considered to be the reason.

In order to clarify the result of the first testrun,a second test run was performed.

In the second test run all four vials showed homogeneous break through times (44:05, 47:27, 47:33 and 48:04 [min:sec])

in the range of two vials of the first test run (43:42 and 43.47 [min:sec]).

The mean breakthrough time of the test substance determined in the second test run Corrositex®assay

was 46 minutes and 47 seconds.

Thus, the results of vial 1 and 2 of the 1sttest run are considered not to be valid and are excluded from evaluation.

The mean breakthrough time of the testsubstance,considering the valid vials of both testruns (6 vials) of the

Corrositex®assay, was 45 minutes and 16seconds.