N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From March 2, 2010 to March 5, 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Human Skin Model Test)

Version / remarks:

adopted 13 April 2004

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

EC Number:

219-145-8

EC Name:

N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine

Cas Number:

2372-82-9

Molecular formula:

C18H41N3

IUPAC Name:

bis(3-aminopropyl)(dodecyl)amine

Test material form:

liquid

Details on test material:

Description: Hazy colourless liquid
Batch: S-001148
Purity: 90.9%
Test substance storage: At room temperature in the dark under nitrogen
Stability under storage conditions: Stable
Expiry date: 04 February 2018

Study specific test substance information:
Test substance handling: When the appearance is becoming cloudy, it might have been cooled for some time. In that case it is best to warm the sample (under nitrogen or in closed bottle) up to about 50 °C, until it is clear again. Draw a sample form the clear solution.

Stability at higher temperatures: Yes, maximum temperature 250°C

Specific details on test material used for the study:

Identification: Dodecyldipropylenetriamine
Chemical name: N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine
Molecular formula: C18H41N3
Molecular weight: 299.54
Description: Hazy colourless liquid (determined at NOTOX)
Batch: S-001148
Purity: 90.9%
Test substance storage: At room temperature in the dark under nitrogen
Stability under storage conditions: Stable
Expiry date: 04 February 2018

Study specific test substance information:
Test substance handling: When the appearance is becoming cloudy, it might have been cooled for some time. In that case it is best to warm the sample (under nitrogen or in closed bottle) up to about 50 °C, until it is clear again. Draw a sample form the clear solution.

Stability at higher temperatures: Yes, maximum temperature 250°C

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Justification for test system used:

Recommended test system in international guidelines (OECD and EC)

Vehicle:

water

Details on test system:

Test system
EpiDerm Skin Model (EPI-200, Lot no.: 12935 kit H): 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.

Source:
MatTek Corporation, Ashland MA, U.S.A.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

Undiluted (50 μl) directly on top of the tissue

Duration of treatment / exposure:

3 minutes and 1 h

Number of replicates:

2 for a 3 minute and 1 h exposure to test substance

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 min exposure

Value:

43

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 h exposure

Value:

42

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Test substance was checked for possible direct MTT reduction by adding the test substance to MTT medium. Because a colour change was observed it was concluded that test substance did interact with MTT. In addition to the normal 1-hour procedure, one freeze-killed tissue treated with test substance and one freeze-killed non treated tissue was used for the cytotoxicity evaluation with MTT. The non-specific reduction of MTT by test substance was 4.72% of the negative control tissues. The net OD of the treated freeze-killed tissues was subtracted from the ODs of the test substance treated viable tissues. The mean absorption at 540 nm measured after treatment with test substance and controls are presented in Table 1. The individual OD540 measurements are presented in Appendix I. Table 2 shows the mean tissue viability obtained after 3-minute and 1-hour treatments with test substance compared to the negative control tissues. Skin corrosion is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after the 3-minute and 1-hour treatments with test substance compared to the negative control tissues was 43% and 42% respectively.

Table 1: Mean absorption in the in vitro skin corrosion test with test substance

	3 minute application				1 hour application
	A (OD540)	B (OD540)	Mean (OD540)	SD	A (OD540)	B (OD540)	Mean (OD540)	SD
Negative control	1.683	1.699	1.691	±0.011	1.707	1.710	1.708	±0.002
Dodecyldipropyl enetriamine	0.702	0.736	0.719	±0.025	0.721	0.716	0.719	±0.004
Positive control	0.150	0.143	0.147	±0.005	0.148	0.145	0.147	±0.002

SD = Standard deviation

Duplicate exposures are indicated by A and B.

(1) The values are corrected for the non-specific MTT reaction.

In this table the values are corrected for background absorption. Isopropanol was used to measure the background absorption

Table 2: Mean tissue viability in the in vitro skin corrosion test with Dodecyldipropylenetriamine

	3 minute application viability (% of control)	1 h application viability (% of control)
Negative control	100	100
Dodecyldipropyl enetriamine	43	42
Positive control	9	9

APPENDIX I: Individual OD measurements at 540 NM

	3 minute application (OD540)		1 hour application (OD540)		1 hour application freeze killed (OD540)
	A	B	A	B	treated	non-treated
Negative control
(OD540) measurement 1	1.686	1.686	1.704	1.713
(OD540) measurement 2	1.694	1.703	1.709	1.714
(OD540) measurement 3	1.670	1.709	1.708	1.702
Dodecyldipropyl enetriamine
(OD540) measurement 1	0.789	0.815	0.820	0.796	0.453	0.373
(OD540) measurement 2	0.779	0.815	0.794	0.798	0.446	0.368
(OD540) measurement 3	0.779	0.815	0.792	0.797	0.444	0.360
Positive control
(OD540) measurement 1	0.155	0.144	0.149	0.146
(OD540) measurement 2	0.149	0.143	0.148	0.144
(OD540) measurement 3	0.146	0.142	0.147	0.145

Applicant's summary and conclusion

Interpretation of results:

Category 1 (corrosive) based on GHS criteria

Conclusions:

Under the study conditions, the test substance was identified as corrosive to skin.

Executive summary:

A study was conducted to determine the skin corrosive property of the test substance (90.9% purity)in vitroaccording to OECD Guideline 431, in compliance with GLP. The test substance was applied undiluted (50 μL) directly on top of the skin tissue. Test substance did interact with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT). In addition to the normal 1 h procedure, one freeze-killed tissue treated with test substance and one freeze-killed non treated tissue was used for the cytotoxicity evaluation with MTT. The non-specific reduction of MTT by test substance was 4.72% of the negative control tissues. The net OD of the treated freeze-killed tissues was subtracted from the ODs of the test substance treated viable tissues. Skin corrosion is expressed as the remaining cell viability after exposure to the test substance. The relative mean tissue viability obtained after 3 min and 1 h treatments with test substance compared to the negative control tissues was 43 and 42%, respectively. Because the mean relative tissue viability for test substance was below 50% after the 3 min treatment, it was considered to be corrosive. Under the study conditions, the test substance was identified as corrosive to skin (Buskens, 2010).