Formaldehyde, reaction products with ethylenediamine

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Remarks:

COMET assay

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

COMET assay agreed as further study following ECHA discussions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 489 (In vivo Mammalian Alkaline Comet Assay)

Version / remarks:

2016

GLP compliance:

yes (incl. QA statement)

Type of assay:

mammalian comet assay

Test material

Test material information

Constituent 1

Constituent 2

Reference substance name:

Schiff bases, C11-14-tert-alkyl methylene

EC Number:

271-766-3

EC Name:

Schiff bases, C11-14-tert-alkyl methylene

Cas Number:

68607-67-0

Molecular formula:

C14H29N

IUPAC Name:

Schiff bases, C11-14-tert-alkyl methylen

Test material form:

liquid

Specific details on test material used for the study:

Identification: Formaldehyde, reaction products with ethylenediamine
CAS Number: 84066-92-2
Physical state/Appearance: Clear colorless liquid
Batch: LC-354-124
Purity: 93% (water 65.6%), ie ca 35%
Expiry Date: 01 January 2019
Storage Conditions: Room temperature in the dark
Intended use/Application: Hydrogen sulphide scavenger

Test animals

Species:

rat

Strain:

Wistar

Details on species / strain selection:

Wistar Han™ (HsdRCCHan™WIST)

Sex:

male

Details on test animals or test system and environmental conditions:

The animals were housed in groups of up to five in solid-floor polypropylene cages with woodflake bedding. Free access to mains drinking water and food (Envigo Teklad 2014 Rodent Pelleted Diet) was allowed throughout the study.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

Water

Details on exposure:

Double dose of maximum 660 mg/kg and killed 4 hours after second dose.

Duration of treatment / exposure:

Two doses,
Sampling 4 hours after second dose

Frequency of treatment:

All animals were dosed twice 24 hours apart

Post exposure period:

4 hours

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

165 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

330 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

660 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Seven

Control animals:

yes, concurrent vehicle

other: Positive control group

Positive control(s):

N-Nitroso-N-methylurea (MNU)

Examinations

Tissues and cell types examined:

Samples of liver, glandular stomach, duodenum and gonadal somatic cells (testes) were obtained from each animal.
Bone marrow was also sampled from the animals of the vehicle control group and the MTD dose group for slide preparation.

Details of tissue and slide preparation:

Sub-samples of the liver, glandular stomach, duodenum and gonadal somatic cells (testes) were taken from the vehicle control animals and the dose group animals and preserved in 10% buffered formalin for possible histopathology investigations. Assessment of cytotoxicity by histopathology is conducted if the results from the Comet assay, or other observations, suggest cytotoxicity may be confounding the interpretation of the Comet assay.

Results and discussion

Test results

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

In the range finding test, there was one death at 880 mg/kg, so the top dose of 660 mg/kg was used with hunched posture, lethargy, ataxia, ptosis and pilo-erection with significant weight loss between the start of the test and its death. Necropsy revealed hemorrhaging along the limiting ridge of the stomach.

Applicant's summary and conclusion

Conclusions:

The test item, Formaldehyde, reaction products with ethylenediamine did not induce any increases in the percentage (comet) tail intensity or median percentage tail intensity in the liver, glandular stomach, duodenum or gonadal somatic cells (testes) and therefore the test item was considered to be unable to induce DNA strand breakage to these tissues in vivo, under the conditions of the test.