3-aminopropyldiethylamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

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:

1988

Report date:

1988

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-aminopropyldiethylamine

EC Number:

203-236-4

EC Name:

3-aminopropyldiethylamine

Cas Number:

104-78-9

Molecular formula:

C7H18N2

IUPAC Name:

(3-aminopropyl)diethylamine

Details on test material:

- Name of test material (as cited in study report): Diethylaminopropylamine
- Physical state: liquid
- Analytical purity: 99.83%
- Purity test date: 25/04/1988
- Lot/batch No.: 8804193
- Expiration date of the lot/batch: no data
- Storage condition of test material: no data

Method

Target gene:

Histidine operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100, TA 1538

Metabolic activation:

with and without

Metabolic activation system:

Liver S9 mix from rats induced with PCB (Pentachlorobiphenyle)

Test concentrations with justification for top dose:

50, 100, 500, 750, 1000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

Details on test system and experimental conditions:

DETERMINATION OF CYTOTOXICITY (preliminary range-finding test)
- Test: in TA 98 and TA 100 strains, with or without S9 mix ; 4 dose-levels (three plates/dose level): 100, 500, 750, 1000, 5000 and 10000µg/plate
- Method: relative total growth (decrease in the number of revertant colonies and/or a thinning of the bacterial lawn);

EXPERIMENTS
Number of independent experiments: 2.

METHOD OF APPLICATION:
* Preincubation:

DURATION
- Preincubation period: 20 min 37°C
- Exposure duration: 48H

NUMBER OF REPLICATIONS: triplicates for all tested doses. Duplicates for positive controls.

CONTROLS
* Without S9 mix
- 5 µg/plate Sodium azide (NAN3) for TA 1535; TA 100 strains
- 100 µg/plate 9-Aminoacridine (9AA) for TA 1537 strain
- 5 µg/plate 2-Nitrofluorene (2NF) for TA 98 strain and TA 1538

* With S9 mix:
- 5 µg/plate 2-Aminoacridine (2AA) for all strains

Evaluation criteria:

Reproducible increase in the number of revertant colonies (2-fold for TA98, TA100, TA 1535 and TA 1537) compared with vehicle controls at any dose-level and/or evidence of a dose-relationship.
Reference to historical data and consideration to biological relevance may also be taken into account.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 750 AND 1000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 750 and 1000µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Test results 3

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 750 AND 1000µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Test results 4

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 750 and 1000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Test results 5

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 750 and 1000µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING STUDY:
- Results on cytotoxicity: Cytotoxicity has been highlighted at 750µg/plate and above.
- Selection of doses for experiment: 50, 100, 500, 750, 1000µg/plate

CYTOTOXICITY: during the main study, cytotoxicty has been observed at 750 and 1000µg/plate (same as in the range-findings study)

Applicant's summary and conclusion

Conclusions:

DEAPA did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.

Executive summary:

The potential of 3 -aminopropyldiethylamine (DEAPA) to induce reverse mutation in Salmonella typhimurium (strains: TA 1535, TA 1537, TA 98, TA 100 and TA 1538) was evaluated in accordance with the international guidelines (OECD 471, Commission Directive No. B13/14) in compliance with the Principles of Good Laboratory Practice. The test item was tested in two independent experiments, with and without a metabolic activation system, both performed according to the preincubation method (20 min at 37°C). Bacterias were exposed to the test item at five dose-levels (three plates/dose-level) selected from a preliminary toxicity test: 50, 100, 500, 750 and 1000 µg/plate. After 48 hours of incubation at 37°C, the revertant colonies were scored. The test item did not induce any noteworthy increase in the number of revertants, both with and without S9 mix, in any of the five strains. Under these experimental conditions, DEAPA did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.