2-[(3-aminopropyl)methylamino]ethanol

Endpoint summary

Description of key information

Skin corrosion/irritation:

In a K1 in vitro skin corrosion test (according to OECD guideline 431; Envigo, 2018), the test substance was found to be corrosive to the skin based on the criteria of the CLP regulation (EC) No 1272/2008.

Eye irritation:

No in vitro or in vivo eye irritation testing is required with the test substance as the substance is demonstrated to be corrosive to the skin.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-05-09 to 2018-05-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Version / remarks:

29 July 2016

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Method B.40 bis of Commission Regulation (EC) No 440/2008

Version / remarks:

30 May 2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch: PFW110370
Purity: 99.10%

Storage Conditions: Room temperature in the dark

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

other: Cell type: epithelial, derived from human skin, and formed into a stratified, cornified epithelium

Source strain:

not specified

Details on animal used as source of test system:

not applicable

Justification for test system used:

This model incorporates several features, which make it advantageous in the study of potential dermal corrosivity.

Vehicle:

unchanged (no vehicle)

Details on test system:

RECONSTRUCTED HUMAN EPIDERMIS (RHE) TISSUE
- Model used: EpiDerm™ Reconstructed Human Epidermis
- Tissue batch number(s): 28607
- Production date: not specified
- Shipping date: not specified
- Delivery date: 08 May 2018
- Date of initiation of testing: 09 May 2018

TEMPERATURE USED FOR TEST SYSTEM
- Temperature used during treatment / exposure: 37 °C
- Temperature of post-treatment incubation (if applicable): 37 °C

REMOVAL OF TEST MATERIAL AND CONTROLS
- Volume and number of washing steps: Rinsing was achieved by filling and emptying each tissue under a constant soft stream of Dulbecco’s Phosphate Buffered Saline (DPBS) (without Ca++ Mg++) for approximately 40 seconds
- Observable damage in the tissue due to washing: n/a
- Modifications to validated SOP: n/a

MTT DYE USED TO MEASURE TISSUE VIABILITY AFTER TREATMENT / EXPOSURE
- MTT concentration: 1.0 mg/mL solution
- Incubation time: 3 Minute and 60 Minute exposure periods
- Spectrophotometer: Labtech LT 4500 microplate reader
- Wavelength: 570 nm (OD570)

NUMBER OF REPLICATE TISSUES: duplicates

CONTROL TISSUES USED IN CASE OF MTT DIRECT INTERFERENCE
- Fresh tissues / killed tissues : Freeze-killed
- Procedure used to prepare the killed tissues (if applicable): Placing untreated EPIDERMTM tissues in an empty 12 well plate and storing in a freezer (-14 to -30 °C) for a minimum of 24 hours. Before use each tissue was thawed by placing in 0.9 mL of assay medium for approximately 1 hour at room temperature.
- No. of replicates per exposure: Two

DECISION CRITERIA
- The test substance is considered to be corrosive to skin if the viability after 3 minutes exposure is less than 50%, or if the viability after 3 minutes exposure is greater than or equal to 50 % and the viability after 1 hour exposure is less than 15%.
- The test substance is considered to be non-corrosive to skin if the viability after 3 minutes exposure is greater than or equal to 50% and the viability after 1 hour exposure is greater than or equal to 15%.

Control samples:

yes, concurrent negative control

Amount/concentration applied:

TEST MATERIAL
- Amount(s) applied: 50 µL of the test item

VEHICLE : no vehicle used

NEGATIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL of sterile distilled water (negative control)

POSITIVE CONTROL
- Amount(s) applied (volume or weight): 50 µL of 8.0 N Potassium Hydroxide (positive control)

Duration of treatment / exposure:

3 Minute exposure and 60 Minute exposure

Duration of post-treatment incubation (if applicable):

n/a

Number of replicates:

Two at each timepoint

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Main test - 60 minute exposure

Value:

9

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Main test - 3 minute exposure

Value:

60.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

- OTHER EFFECTS:
- Visible damage on test system: not observed
- Direct-MTT reduction: The MTT solution containing the test item turned purple, indicating that the test item was able to directly reduce MTT. An additional procedure using freeze killed tissues was performed to determine the magnitude of interference due to direct reduction of MTT. However, the results obtained showed that the magnitude of interference was negligible. It was therefore considered unnecessary to use the results of the freeze-killed tissues for quantitative correction of results or reporting purposes.
- Colour interference with MTT: By visual assessment, the solution containing the test item did not become colored. This was taken to indicate the test item did not have the potential to cause color interference.

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes, the mean OD570 for the negative control treated tissues was 1.755 for the 3-Minute exposure period and 1.919 for the 60-Minute exposure period.
- Acceptance criteria met for positive control: yes, the relative mean tissue viability for the positive control treated tissues was 2.8% relative to the negative control following the 60-Minute exposure period.
- Acceptance criteria met for variability between replicate measurements: yes, the viability Coefficient of Variation between the two tissue replicates were 1.6% (3 minutes); 0.9% (60 minutes) for the negative control and 16.7% (3 minutes) for the test item. The remainder of the Coefficient of Variations were not applicable because the viabilities were < 20%.

Mean OD₅₇₀Values and Viabilities for the Negative Control Item, Positive Control Item and Test Item

Tissue	Exposure Period	MeanOD570of individual tissues	Mean OD570of duplicate tissues	Standard Deviation	Coefficient of Variation (%)	Relative Mean Viability (%)
Negative Control	3 Minutes	1.735	1.755	0.028	1.6	100*
		1.774
	60 Minutes	1.906	1.919	0.018	0.9
		1.931
Positive Control	3 Minutes	0.064	0.060	0.006	na	3.4
		0.055
	60 Minutes	0.056	0.054	0.003	na	2.8
		0.052
Test Item	3 Minutes	1.190	1.065	0.177	16.7	60.7
		0.939
	60 Minutes	0.160	0.172	0.017	na	9.0
		0.184

OD= Optical density

*=          The mean percentage viability of the negative control tissue is set at 100%

na=         Not applicable

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

The test item was considered to be corrosive. According to UN GHS, it is classified as H314 (Combination of sub-categories 1B and 1C).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance is classified as skin corrosion, leading to classification as serious eye damage (Category 1)

Endpoint conclusion

Endpoint conclusion:

no study available

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation/corrosion:

An in vitro EPIDERM(TM) Skin Corrosion Test was performed according to OECD guideline 431 (Envigo, 2018). The test substance is applied neat on human non-transformed keratinocytes for 3 mins or 1 hr. The tissue viability after exposure was determined to be 60.7% after 3 mins, and 9% after 1hr. The substance is therefore considered to be corrosive to the skin.

Eye irritation:

An in vitro eye irritation study was waived based on the justification that the test item is classified as skin corrosive, leading to a classification as serious eye damage category 1.

Justification for classification or non-classification

Skin irritation/corrosion: According to the in vitro skin corrosion study, the test substance is corrosive to the skin and therefore classified as category 1B according to the criteria of the CLP Regulation (EC) No 1272/2008.

Eye irritation: Based on the classification of the test item as skin corrosive (category 1B), the test item is classified for serious eye damage category 1 according to the CLP Regulation (EC) No 1272/2008.