2,2'-dimorpholinyldiethyl ether

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

Remarks:

Guideline study, GLP study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

(only 2-aminoanthracene as positive control with metabolic activation)

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:

2,2'-dimorpholinyldiethyl ether

EC Number:

229-194-7

EC Name:

2,2'-dimorpholinyldiethyl ether

Cas Number:

6425-39-4

Molecular formula:

C12H24N2O3

IUPAC Name:

4-{2-[2-(morpholin-4-yl)ethoxy]ethyl}morpholine

Test material form:

liquid

Details on test material:

- State of aggregation : liquid
- Appearance : colourless to yellow brown

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: H 50541 H x B (Substance number: 95/452)
- Analytical purity: 98.4%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature (N2 conditions)
- Stability under test conditions: guaranteed throughout study period via reanalysis

OTHER SPECIFICS:
- Name of test substance (as cited in study report): 2,2' -Dimorpholinodiethylether
- Physical state: liquid

Method

Target gene:

His- and Trp-Operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9-mix

Test concentrations with justification for top dose:

0, 20, 100, 500, 2,500, 5,000 µg/plate (standard plate test and preincubation test)

Vehicle / solvent:

water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: with S9: 2-aminoanthracene (all tester strains); without S9: N-methyl-N'-nitro-N-nitrosoguanidine (TA 1535, TA 100), 4-nitro-o-phenylendiamine (TA 98), 9-aminoacridine (TA 1537), N-ethyl-N'-nitro-N-nitrosoguanidine (E. coli WP2 uvrA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION (preincubation test)
- Preincubation period: 20 min
- Exposure duration: 48 - 72 h

DURATION (standard plate test)
- Exposure duration: 48 - 72 h

NUMBER OF REPLICATIONS: 3 plates/dose/experiment.


DETERMINATION OF CYTOTOXICITY
- Method: reduction of the number of revertants compared to negative control; reduction of titer; reduction of backgroud growth

Evaluation criteria:

The test subtance is considered positive in this assay if the following criteria are met: A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S9-mix or after adding a metabolizing system.

Results and discussion

Test results

Test results 1

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:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: no precipitation up to 5,000 µg/plate (highest dose tested)

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test:

Dose (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli WP2 uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	20±5	19±2	128±10	123±4	10±1	10±1	31±4	47±5	32±1	42±2
20	19±2	18±2	115±20	125±12	10±2	11±4	32±3	46±4	33±5	41±3
100	19±5	17±3	111±7	121±14	10±1	7±1	31±5	39±8	30±2	37±6
500	19±3	14±1	128±8	107±14	9±3	8±1	27±5	33±10	30±1	39±2
2500	19±3	16±4	123±7	121±15	10±1	6±0	31±4	38±5	32±4	42±2
5000	18±3	15±2	140±3	97±17	8±1	8±1	28±6	28±8	30±3	43±2
2 -AA	-	149±7	-	1467± 133	-	149±24	-	1241± 29	-	236± 17
MNNG	783± 37	-	1027± 49	-	-	-	-	-	-	-
AAC	-	-	-	-	804±13	-	-	-	-	-
NOPD	-	-	-	-	-	-	1308± 135	-	-	-
ENNG	-	-	-	-	-	-	-	-	521± 26	-

Mean ± SD

 

Preincubation-test:

Dose (µg/plate)	TA1535		TA100		TA1537		TA98		E. coli WP2 uvrA
	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
0	18±1	19±2	133±4	130±17	11±2	11±3	29±1	40±3	32±4	38±3
20	18±2	16±2	116±9	108±9	11±1	11±2	23±2	42±1	31±6	32±5
100	21±1	17±0	124±6	121±10	8±1	11±1	24±2	36±5	28±4	33±5
500	17±2	14±1	111±0	125±13	8±2	11±2	24±2	38±1	30±6	25±4
2500	19±1	16±3	117±15	132±7	8±2	9±1	25±1	40±3	28±3	28±4
5000	16±2	15±2	102±16	138±14	7±1	7±2	20±3	38±7	26±2	30±51
2-AA	-	116±13	-	699± 2	-	119±9	-	727± 12	-	177±11
MNNG	978± 135	-	1177± 126	-	-	-	-	-	-	-
AAC	-	-	-	-	402±62	-	-	-	-	-
NOPD	-	-	-	-	-	-	1148± 103	-	-	-
ENNG	-	-	-	-	-	-	-	-	569±49	-

Mean ± SD

X: reduced background growth

2-AA: 2-aminoanthracene

MNNG; N-methyl-N-nitro-N-nitrosoguanidine

ENNG; N-ethyl-N-nitro-N-nitrosoguanidine

NOPD: 4-nitro-o-phenylendiamine

AAC: 9-aminoacridine chloride monohydrate

 

According to the results of the study, the test substance is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here. The positive controls gave the expected results.

 

Applicant's summary and conclusion

Conclusions:

According to the results of the study, the test substance is not mutagenic in the Ames test and in the Escherichia coli - reverse mutation assay under the experimental conditions chosen here. The positive controls gave the expected results.