2-dibutylaminoethanol

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline-Study with acceptable restrictions (original study report in japanese, english translation available).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(adopted on July 21, 1997)

Deviations:

yes

Remarks:

(2-Aminoanthracene was used as sole positive control in tests with S9 mix)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2-dibutylaminoethanol

EC Number:

203-057-1

EC Name:

2-dibutylaminoethanol

Cas Number:

102-81-8

Molecular formula:

C10H23NO

IUPAC Name:

2-(dibutylamino)ethan-1-ol

Details on test material:

- Name of test material (as cited in study report): 2-(Di-n-butylamino)ethanol
- Physical state: yellow, transparent liquid
- Analytical purity: 99.8%
- Impurities (identity and concentrations): 0.12% water as impurity
- Storage condition of test material: Kept protected from light at room temperature

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:

S9: rat liver, induced with phenobarbital and 5,6-benzoflavone

Test concentrations with justification for top dose:

-S9 mix; 0, 156, 313, 625, 1250, 2500, 5000 μg/plate (all strains)
+S9 mix; 0, 19.5, 39.1, 78.1, 156, 313, 625 μg/plate (all strains)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: test substance was insoluble in water

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9 mix: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF2M; in DMSO; TA100, TA98, WP2 uvrA), sodium azide (SA; TA1535) and 9-aminoacridine (9AA; in DMSO; TA1537); +S9 mix; 2-Aminoanthracene (2AA; in DMSO; all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 plates; the test was conducted twice with regard to same dose

DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition

OTHER EXAMINATIONS: 0.1 mL of the test substance solution of highest dose and 0.5 mL of S9 mix were separately dropped onto the culture plate to check mixing-in of other bacteria at the end of culture.

OTHER: The number of revertant colonies produced was counted by a colony analyzer (System Science Co., CA-11) or visually.

Evaluation criteria:

If the median of the number of revertant colonies in the plate containing the test substance from one sort or more strains out of 5 strains in total in the absence of S9 mix or in the presence of S9 mix increased twice or more than that of the negative control value and its increase had reproducibility and dose dependency, it shall be judged that the said bacteria has mutagenicity (positive) in the present test system.

Statistics:

No data.

Results and discussion

Test results

Test results 1

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Growth inhibition was observed at and above 2500 μg/plate without S9 mix, and at 625 μg/plate (highest dose) with S9 mix in all four S. typhimurium strains.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

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:

cytotoxicity

Remarks:

Growth inhibition was observed without S9 mix at 5000 μg/plate (highest dose) and at 625 μg/plate (highest dose) in the presence of S9 mix.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation observed.

RANGE-FINDING/SCREENING STUDIES: conducted (concentrations: 50.0, 150, 500, 1500 and 5000 μg/plate), each 3 plates for negative and positive controls, and 1 plate for each dose were used; as a result, the grow inhibition was observed in the highest dose of 5000 μg/plate for all the test bacteria in the absence of S9 mix and in the doses of 500 μg/plate and more for all the test bacteria in the presence of S9 mix. In addition, precipitations from the test substance were not observed in all the doses used. From the said results, a dose of 5000 μg/plate was determined for all the test bacteria in the absence of S9 mix and a dose of 625 μg/plate was determined for all the test bacteria in the presence of S9 mix.

ADDITIONAL INFORMATION ON CYTOTOXICITY: As a result, the grow inhibition was observed in a dose of 2500 μg/plate or more in the test bacteria TA100, TA1535, TA98 and TA1537 in the absence of S9 mix, in the highest dose of 5000 μg/plate in WP2 uvrA and in a dose of 625 μg/plate in all the test bacteria in the presence of S9 mix.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Test results of experiment 1 (preincubation test).

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	E. coli WP2 uvrA	TA98	TA1537
–	0	142 ± 10	9 ± 1	21 ± 3	25 ± 3	7 ± 2
–	156	136 ± 1	7 ± 2	24 ± 5	26 ± 3	8 ± 1
–	313	145 ± 40	7 ± 2	26 ± 3	24 ± 4	7 ± 1
–	625	163 ± 10	7 ± 1	24 ± 6	29 ± 1	10 ± 1
–	1250	150 ± 11	6 ± 4	22 ± 1	27 ± 5	9 ± 3
–	2500	163* ± 11	7* ± 3	23 ± 8	26* ± 5	7* ± 2
–	5000	0* ± 0	0* ± 0	0* ± 0	0* ± 0	0* ± 0
Positive controls, –S9	Name	AF2	SA	AF2	AF2	9AA
	Concentrations (μg/plate)	0.01	0.5	0.01	0.1	80
	Mean No. of colonies/plate (average of 3 ± SD)	602 ± 24	627 ± 18	178 ± 12	380 ± 48	226 ± 4
+	0	160 ± 12	10 ± 4	34 ± 3	40 ± 3	14 ± 2
+	19.5	149 ± 22	8 ± 0	26 ± 3	35 ± 9	12 ± 1
+	39.1	163 ± 4	10 ± 2	28 ± 2	35 ± 4	14 ± 2
+	78.1	176 ± 29	7 ± 1	28 ± 6	35 ± 6	15 ± 3
+	156	162 ± 13	11 ± 3	25 ± 4	35 ± 2	12 ± 4
+	313	162 ± 9	9 ± 5	31 ± 2	34 ± 10	13 ± 4
+	625	151* ± 14	9* ± 5	29* ± 1	47* ± 9	11* ± 1
Positive controls, +S9	Name	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1	2	10	0.5	2
	Mean No. of colonies/plate (average of 3 ± SD)	821 ± 22	338 ± 24	796 ± 60	441 ± 46	268 ± 8

*growth inhibition was observed

AF2: 2-(2-FuryI)-3-(5-nitro-2-furyl)acrylamide, SA: Sodium azide, 9AA: 9-Aminoacridine, 2AA: 2-Aminoanthracene

 

Table 2. Test results of experiment 2 (preincubation test).

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	E. coli WP2 uvrA	TA98	TA1537
–	0	118 ± 12	8 ± 3	41 ± 5	23 ± 5	12 ± 3
–	156	117 ± 12	9 ± 4	42 ± 8	24 ± 5	8 ± 1
–	313	124 ± 17	8 ± 3	53± 9	23 ± 5	12 ± 3
–	625	125 ± 8	8 ± 3	42 ± 7	23 ± 4	8 ± 2
–	1250	123 ± 2	9 ± 2	38 ± 3	23 ± 4	6 ± 3
–	2500	137* ± 4	7* ± 2	42 ± 10	20* ± 7	7* ± 3
–	5000	0* ± 0	0* ± 0	0* ± 0	0* ± 0	0* ± 0
Positive controls, –S9	Name	AF2	SA	AF2	AF2	9AA
	Concentrations (μg/plate)	0.01	0.5	0.01	0.1	80
	Mean No. of colonies/plate (average of 3 ± SD)	431 ± 47	601 ± 1	176 ± 21	422 ± 38	313 ± 97
+	0	128 ± 7	8 ± 3	46 ± 4	40 ± 4	13 ± 1
+	19.5	132 ± 9	8 ± 3	44 ± 5	35 ± 3	12 ± 6
+	39.1	131 ± 23	8 ± 3	53 ± 5	31 ± 5	14 ± 4
+	78.1	130 ± 6	6 ± 3	44 ± 4	38 ± 3	13 ± 5
+	156	124 ± 8	9 ± 2	49 ± 5	38 ± 1	13 ± 2
+	313	142 ± 9	5 ± 2	46 ± 3	33 ± 6	12 ± 5
+	625	132* ± 6	7* ± 1	48* ± 3	35* ± 6	14* ± 2
Positive controls, +S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	1	2	10	0.5	2
	Mean No. of colonies/plate (average of 3 ± SD)	770 ± 27	345 ± 24	777 ± 57	434 ± 14	253 ± 20

*growth inhibition was observed

AF2: 2-(2-FuryI)-3-(5-nitro-2-furyl)acrylamide, SA: Sodium azide, 9AA: 9-Aminoacridine, 2AA: 2-Aminoanthracene