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Toxicological information

Genetic toxicity: in vitro

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Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP-guideline study, tested with the source substance 2-amino-2-ethyl-1,3-propanediol (CAS 115-70-8). In accordance to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Justification for type of information:
The justification for category approach/read-across has been attached in section 13 of this IUCLID. In the enclosed document arguments are given for a category approach for four 2-amino-1,3-propane-diols. These substances share a common propane backbone with an amine group at 2-carbon position and primary alcohols at 1 and 3 positions. The members of the aminopropanediol category are: 2-amino-1,3-propanediol (APD, CAS No. 534-03-2), 2-amino-2-methyl-1,3-propane-diol (AMPD, CAS No. 115-69-5), 2-amino-2-ethyl-1,3-propanediol (AEPD, CAS No. 115-70-8), and 2-amino-2-(hydroxymethyl)-1,3-propanediol (trometamol, CAS No. 77-86-1).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2004
Report Date:
2004

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Qualifier:
according to
Guideline:
JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): 2-amino-2-ethyl-1,3-propanediol
- Analytical purity: 99.4%

Method

Target gene:
his operon (for S. typhimurium strains)
trp operon (for E. coli strains)
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
cofactor supplemented post-mitochondrial fraction (S9-mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone
Test concentrations with justification for top dose:
156, 313, 625, 1250, 2500 and 5000 μg/plate with and without metabolic activation
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: water
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: without S9-mix
Remarks:
- S9-mix: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (0.01 µg/plate) for TA100 and E.coli, (0.1 µg/plate) for TA 98; sodium azide (0.5 µg/plate for TA1535); 2-methoxy-6-chloro-9-[3-(2-chloroethyl)- aminopropylamino] acridine-2HCl for (1.0 µg/plate) TA1537
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: with S9-mix
Remarks:
+ S9-mix: Benzo[a]pyrene (5.0 µg/plate) for TA100, TA98 and TA1537; 2-aminoanthracene (2.0 µg/plate) for TA1535 and (10.0 µg/plate) for E.coli WP2 uvrA
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 for each concentration

DETERMINATION OF CYTOTOXICITY
- Method: The inhibition of growth of the test bacterial strains by the test substance was observed with a stereoscopic microscope.

Evaluation criteria:
When the number of reverse mutation colonies increased by almost twice the solvent control or more, and reproducibilityor dependence on the dose of the test substance was observed, the result was considered positive.

Results and discussion

Test resultsopen allclose all
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:
not examined
Positive controls validity:
valid
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:
not examined
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: Precipitation of a white crystalline substance was observed in the test substance treatment groups of 2500 µg/plate or higher in the presence of the metabolic activation system, regardless of the kind of bacterial strain used.

RANGE-FINDING/SCREENING STUDIES:
In the range finding study, 8 treatment doses of the test solution were used in the range of 0.305 - 5000 µg/plate. Precipitation of a white crystalline substance was observed at the highest dose of 5000 µg/plate in the presence of a metabolic activation system, regardless of the kind of bacterial strain used. No inhibition of growth of bacterial strains was observed, regardless of the kind of strain and the presence or absence of metabolic activation.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
No inhibition of growth of bacterial strains was observed, regardless of the kind of strain and the presence or absence of metabolic activation (data not shown).
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Results of the main 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

TA 1535

E.coli WP2 uvrA

TA 98

TA 1537

-

0

113 ± 21.4

10 ± 1.0

29 ± 3.5

17 ± 3.1

12 ± 1.5

-

156

110 ± 12.4

8 ± 1.2

28 ± 2.6

12 ± 2.1

9 ± 1.2

-

313

108 ± 13.1

8 ± 2.3

26 ± 3.8

13 ± 4.0

13 ± 2.0

-

625

107 ± 4.7

7 ± 1.0

27 ± 0.6

16 ± 1.7

10 ± 1.0

-

1250

114 ± 7.1

7 ± 2.1

27 ± 4.6

13 ± 3.1

10 ± 2.1

-

2500

115 ± 2.3

9 ± 4.2

28 ± 4.4

15 ± 3.0

10 ± 2.0

-

5000

114 ± 7.4

7 ± 1.7

19 ± 3.1

12 ± 2.5

10 ± 1.7

Positive controls,

-S9

Name

 

Concentrations (µg/plate)

 

Mean No. of colonies/plate (average of 3 ± SD)

AF-2

 

0.01

 

 

469 ± 18.1

SAZ

 

0.50

 

 

176 ± 34.7

AF-2

 

0.01

 

 

235 ± 29.5

AF-2

 

0.10

 

 

363 ± 9.8

ICR-191

 

1.0

 

 

1194 ± 138.0

 

 

TA 100

TA 1535

E.coli WP2 uvrA

TA 98

TA 1537

+

0

126 ± 5.9

13 ± 0.6

29 ± 2.9

20 ± 2.0

12 ± 2.6

+

156

123 ± 2.5

8 ± 0.6

27 ± 2.7

19 ± 0.6

14 ± 1.7

+

313

127 ± 12.2

8 ± 0.6

26 ± 4.4

20 ± 1.5

13 ± 2.0

+

625

106 ± 6.0

9 ± 1.5

25 ± 2.5

19 ± 2.1

11 ± 1.2

+

1250

125 ± 10.2

10 ± 2.1

22 ± 3.1

23 ± 2.1

12 ± 1.5

+

2500

126 ± 3.2

8 ± 2.1

19 ± 2.1

18 ± 2.5

12 ± 3.0

+

5000

135 ± 9.0

7 ± 2.1

21 ± 3.8

14 ± 2.9

10 ± 2.3

Positive controls, +S9

Name

 

Concentrations (µg/plate)

 

Mean No. of colonies/plate (average of 3 ± SD)

B[a]P

 

5.00

 

 

621 ± 23.8

2AA

 

2.00

 

 

117 ± 16.7

2AA

 

10.0

 

 

407 ± 12.1

B[a]P

 

5.00

 

 

247 ± 5.5

B[a]P

 

5.00

 

 

111 ± 6.7

 

SAZ: Sodium azide

AF-2: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide

ICR-191: 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)-aminopropylamino]acridine*2HCl

B[a]P: Benzo[a]pyrene

2AA: 2-Aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative