Formaldehyde, oligomeric reaction products with 4,4'-isopropylidenediphenol and diethylenetriamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes

Test material

Test material information

1

Reference substance name:

Formaldehyde, oligomeric reaction products with 4,4'-isopropylidenediphenol and diethylenetriamine

EC Number:

500-263-6

EC Name:

Formaldehyde, oligomeric reaction products with 4,4'-isopropylidenediphenol and diethylenetriamine

Cas Number:

77138-45-5

Molecular formula:

(C15H16O2.C4H13N3.CH2O)x

IUPAC Name:

2-[({2-[(2-aminoethyl)amino]ethyl}amino)methyl]-4-(2-{3-[({2-[(2-aminoethyl)amino]ethyl}amino)methyl]-4-hydroxyphenyl}propan-2-yl)phenol; 2-[({2-[(2-aminoethyl)amino]ethyl}amino)methyl]-4-[2-(4-hydroxyphenyl)propan-2-yl]phenol; bis(2-aminoethyl)amine

Test material form:

liquid

In vitro test system

Test system:

human skin model

Source species:

human

Vehicle:

unchanged (no vehicle)

Details on test system:

EPISKIN™ Reconstructed Human Epidermis Model Kit
Supplier : MatTek
Date received : 24 May 2016
EpiSkinTM Tissues (0.38cm2) lot number : 23338
Maintenance Medium lot number : 051916TMA

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

50 uL

Duration of treatment / exposure:

3 minutes
60 minutes

Duration of post-treatment incubation (if applicable):

3 hours

Number of replicates:

3

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minutes

Value:

61.6

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

60 minutes

Value:

2.6

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Tissue	Exposure Period	Mean OD562of individual tissues (tvt)	Mean OD562of duplicate tissues	Corrected OD562of tissues (tvt -[tktukt])	Standard Deviation	Coefficient of Variation (%)	Relative Mean Viability (%)
Negative Control⊕	3 Minutes	1.720	1.835		0.162	8.8	100*
		1.949
	60 Minutes	1.872	1.838		0.048	2.6
		1.804
Positive Control⊕	3 Minutes	0.100	0.094		0.009	N/A	5.1
		0.087
	60 Minutes	0.090	0.087		0.005	N/A	4.7
		0.083
Test Item	3 Minutes	1.347	1.415	1.130	0.095	6.7	61.6
		1.482
	60 Minutes	0.562	0.409	0.048	0.216	N/A	2.6
		0.256

3 minute exposure corrected mean OD562= 0.412 (tkt) – 0.127 (ukt) = 0.285

60 minute exposure corrected mean OD562= 0.474 (tkt) – 0.113 (ukt) = 0.361

tvt = treated viable tissues

tkt = treated killed tissues

ukt = untreated killed tissues

Applicant's summary and conclusion

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

The test item was considered to be corrosive to the skin.

Executive summary:

Introduction

The purpose of this test is to evaluate the corrosivity potential of the test item using the EpiDerm™ Human Skin Model after treatment periods of 3 and 60 minutes.

Corrosion is directly related to cytotoxicity in the EpiDerm™ tissue. Cytotoxicity is determined by the reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) to formazan by viable cells in the test item treated tissues relative to the corresponding negative control. The results are used to make a prediction of the corrosivity potential of the test item.

Methods

Duplicate tissues were treated with the test item for exposure periods of 3 and 60 minutes. The test item was found to directly reduce MTT and therefore additional non-viable tissues were incorporated into the testing for correction purposes. Negative and positive control groups were treated for each exposure period. At the end of the exposure period the test item was rinsed from each tissue before each tissue was taken for MTT-loading. After MTT loading each tissue was placed in 2 mL Isopropanol for MTT extraction.

At the end of the formazan extraction period each well was mixed thoroughly and triplicate 200 μL samples were transferred to the appropriate wells of a pre-labeled 96-well plate. The optical density (OD) was measured at 562 nm (OD562).

Data are presented in the form of percentage viability (MTT reduction in the test item treated tissues relative to negative control tissues).

Results

The relative mean viabilities for each treatment group were as follows:

Exposure Period                                         Percentage Viability

Negative Control       Positive Control       Test Item

3 minute                     100*                                   5.1                     61.6

60 minute                    100*                                   4.7                     2.6

*The mean viability of the negative control tissues is set at 100%

Quality criteria: The quality criteria required for acceptance of results in the test were satisfied.

Conclusion

The test item was considered to be corrosive to the skin.