N-(2-aminoethyl)-1,3-propanediamine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-10-18 - 1990-12-12

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Meets generally accepted scientific standards, well documented and acceptable for assessment; only four strains were tested.

Justification for type of information:

Corresponding to the draft decision (CCH-D-2114618994-36-01/D) the registrant agrees to conduct an OECD 471.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Report date:

1990

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

, only four strains

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-(2-aminoethyl)-1,3-propanediamine

EC Number:

236-882-0

EC Name:

N-(2-aminoethyl)-1,3-propanediamine

Cas Number:

13531-52-7

Molecular formula:

C5H15N3

IUPAC Name:

N1-(2-Aminoethyl)-1,3-propanediamine

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

S9-Mix prepared from sprange Dawlay rat livers after Aroclor 1254 activation

Test concentrations with justification for top dose:

0, 20 ,100, 500, 2500, 5000 µg/plate (TA 100, TA 1537, TA 98; SPT)
0, 500, 1000, 5000, 7000 µg/plate (TA 1535; SPT)
0, 4, 20, 100, 500, 2500 µg/plate (all tester strains; PIT)

Vehicle / solvent:

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

Controls

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: see details

Details on test system and experimental conditions:

Standard plate test
Test tubes containing 2 ml portions of saft agar kept in a water bath at 45°C, and the remaining companents are added in the following order:
0.1 mL test solution
0.1 mL bacterial suspension
0.5 mL S-9 mix (in tests with metabolic activatian) or
0.5 mL phosphate buffer (in tests without metabalic activatian)
After mixing, the samples are poured onto Vogel-Bonner agar plates. After incubation at 37°C for 48 -72 hours in the dark, the bacterial calonies (his+ revertants) are counted.


Preincubation test
0.1 mL test solution or vehicle, 0.1 ml bacterial Suspension and 0.5 mL S-9 mix are incubated at 37°C for the duration of 20 minutes. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the agar plates within approx.30 seconds.

Both Tests
In each experiment 3 Test plates per dose per control used. After incubation at 37°C for 48 hours in the dark, the bacterial colonies (his+ revertants) are counted.

Posivite Control:
with S-9 mix:
10 µg 2-aminoanthracene (2-AA) (dissolved in DMSO) for the strains TA 100, TA 98, TA 1537and TA 1535

without S-9 mix:
5 µg N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) (dissolved in DMSO) for the strains TA 100 and TA 1535
10 µg 4-nitro-o-phenylendiamine (NOPD) (dissolved in DMSO) for the strain TA 98
100 µg 9-aminoacridine (AAC) chloride monohydrate (dissolved in DMSO) for the strain TA 1537

The Titer was determined and in regularly measurements the strain characteristics were checked. Sterility control performed.

Evaluation criteria:

In general, a substance to be characterized as positive in the Ames test hat to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Toxicity: A bacteriotoxic effect was observed in the standard plate test with and without S-9 mix at 5000 µg/plate.
In the preincubation assay bacteriotoxicity was found only without metabolic activation at 2500 µg/plate.
Mutagenicity: An increase in the number of his+ revertants was not observed both in the standard plate test and in the preincubation test either without S-9 mix or after the addition of a metabolizing system.

According to the results of the present study, the test substance is not mutagenic in the Ames test under the experimental conditions chosen here. The slightly enhanced figures in the 1st experiment using TA 1535, which might partly be explained by the very 1ow spontaneous rate was not confirmed in two additional standard plate tests and in a preincubation assay. Therefore, these finding are regarded as incidental and not as an indication of a point mutagenic activity.

The positive controls (+S9-mix: 2AA; -S9-mix: MNNG, NOPD, AAC) showed the expected results.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Results in detail (Ames-test, BASF 1990, Project No. 40M0228/894480)

Plate incorporation with S-9 mix (colony number = mean values) - experiment 1:

Dose (µg/per plate)	TA 100	TA 1537	TA 98	TA1535
0 (aqua dest.)	106	11	33	11
20	95	8	34	10
100	96	9	35	13
500	65	11	32	15
2500	72	9	28	26
5000	61	12	21	29
10 µg 2-AA	1563	163	1507	305

Plate incorporation without S-9 mix (colony number = mean values) - experiment 1:

Dose (µg/per plate)	TA 100	TA 1537	TA 98	TA 1535
0 (aqua dest.)	99	6	22	18
20	107	6	22	27
100	112	7	19	25
500	108	8	26	24
2500	95	8	18	23
5000	83	9	-	21
5 µg MNNG	1500			1465
100 µg AAC		539
10 µg NPD			808

Plate incorporation with S-9 mix (colony number = mean values) - experiment 2:

Dose (µg/per plate)	TA 1535
0 (aqua dest.)	18
500	23
1000	22
3000	21
5000	24
7000	19
10 µg AA	195

Plate incorporation without S-9 mix (colony number = mean value) - experiment 2:

Dose (µg/per plate)	TA 1535
0 (aqua dest.)	18
500	27
1000	25
3000	24
5000	23
7000	21
5 µg MNNG	1465

Plate incorporation with S-9 mix (colony number = mean value) - experiment 3:

Dose (µg/per plate)	TA 100	TA 1537	TA 98	TA1535
0 (aqua dest.)	117	15	42	18
4	106	12	45	10
20	120	11	35	12
100	132	11	32	9
500	122	15	24	18
2500	101	10	21	25
10 µg 2-AA	1061	96	687	187

Plate incorporation with S-9 mix (colony number = mean value) - experiment 3:

Dose (µg/per plate)	TA 100	TA 1537	TA 98	TA1535
0 (aqua dest.)	105	12	21	23
4	105	12	14	18
20	120	8	21	19
100	135	12	22	19
500	111	13	21	20
2500	-	2	7	9
5 µg MMNG	981			1010
100 µg AAC		479
10 µg NPD			740

Plate incorporation with S-9 mix (colony number = mean values) - experiment 4:

Dose (µg/per plate)	TA 1535
0 (aqua dest.)	16
500	17
1000	19
3000	24
5000	19
7000	18
10 µg AA	151

Plate incorporation without S-9 mix (colony number = mean value) - experiment 4:

Dose (µg/per plate)	TA 1535
0 (aqua dest.)	14
500	16
1000	19
3000	19
5000	16
7000	10
5 µg MNNG	1777

- : reduced his- background

2-AA: 2-Aminoanthracene (dissolved in DMSO)

MNNG: N-Methyl-N'-nitro-N-nitrosoguanidine (dissolved in DMSO)

AAC: 9-aminoacridine chloride monohydrate (dissolved in DMSO)

NPD: 4-nitro-o-phenylendiamine (dissolved in DMSO)