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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: III. Results From the Testing of 225 Chemicals
Author:
Zeiger E, Anderson B, Haworth S, Lawlor T, Mortelmans K, Speck W
Year:
1987
Bibliographic source:
Environmental Mutagenesis 9 (Suppl 9), 1-110

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
No E.coli strain or TA102 tested
Principles of method if other than guideline:
Haworth S, Lawlor T, Mortelmans K, Speck W, Zeiger E (1983): Salmonella mutagenicity results for 250 chemicals. Environ Mutagen S[Suppl 1]:3-142.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1,1',1''-nitrilotripropan-2-ol
EC Number:
204-528-4
EC Name:
1,1',1''-nitrilotripropan-2-ol
Cas Number:
122-20-3
Molecular formula:
C9H21NO3
IUPAC Name:
1,1',1''-nitrilotripropan-2-ol
Specific details on test material used for the study:
Purity > 94%

Method

Target gene:
His locus
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor-induced male Sprague-Dawley rats and Syrian hamsters S9-mix
Test concentrations with justification for top dose:
0, 100, 333, 1000, 3333, 10000 µg/plate
Vehicle / solvent:
Water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylene diamine: TA 98, without S9-mix; 2-aminoanthracene: all strains, with S9-mix
Details on test system and experimental conditions:
The test chemical (0.05 mL), Salmonella culture (0.10 mL), and S-9 mix or buffer (0.50 mL) were incubated at 37°C, without shaking, for 20 min. The top agar was added and the contents of the tubes were mixed and poured onto the surface of petri dishes containing Vogel-Bonner medium. The histidine-independent (his+) colonies arising on these plates were counted following two days incubation at 37°C. Plates were machine counted unless precipitate was present which interfered with the count, or the color of the test chemical on the plate reduced the contrast between the colonies and the background agar.
Evaluation criteria:
Evaluations were made at both the individual trial and overall chemical levels. Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable (?), or nonmutagenic (-), depending on the magnitude of the increase of his+ revertants, and the shape of the dose-response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of “ +W,” if only a single dose was elevated over the control, or if the increase seen was not dose related. The distinctions between a questionable mutagenic response and a nonmutagenic or weak mutagenic response, and between a weak mutagenic response and mutagenic response are highly subjective. It was not necessary for a response to reach
twofold over background for a chemical to be judged mutagenic. A chemical was judged mutagenic (+) or weakly mutagenic (+ W) if it produced a reproducible dose-related reponse over the solvent control in replicate trials.
A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a “+W” response, or if only single doses produced increases in his+ revertants in repeat trials. Chemicals were judged nonmutagenic (-) if they did not meet the criteria for a mutagenic or questionable response.
Statistics:
no data

Results and discussion

Test results
Key result
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 nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
Triisopropanolame was negative (non-mutagenic) in all assays with and without metabolic activation.

Any other information on results incl. tables

Strain TA1535

Dose

No Activation
(Negative)

10% HLI
(Negative)

10% RLI
(Negative)

Dose units

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

42

3.8

21

2.7

11

1.3

100         

33

1.2

13

1.9

13

0.3

333         

34

1.9

15

1.2

15

2.3

1000         

32

6.4

13

0.3

12

4.6

3333         

24

3.5

16

1.8

11

1

10000         

28

5.5

15

0.3

10

0.9

Positive Control

1162

4.5

115

9.1

99

0.9

 

Strain TA100 

Dose

No Activation
(Negative)

10% HLI
(Negative)

10% RLI
(Negative)

Dose units

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

151

9

141

0.7

151

10.2

100         

141

9

147

4.8

139

0.6

333         

131

0.6

141

5.6

146

2.4

1000         

144

8.6

146

3.2

149

4.3

3333         

152

5

151

10.3

143

7.9

10000         

148

2.5

146

4.7

139

5.8

Positive Control

1406

62.8

1040

19.8

1025

37.8

 

Strain TA98 

Dose

No Activation
(Negative)

10% HLI
(Negative)

10% RLI
(Negative)

Dose units

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

20

1

26

4.5

30

1.9

100         

19

1.8

28

0.9

33

4.9

333         

19

2.9

31

1.5

29

2

1000         

21

0

31

4.1

30

0.9

3333         

15

0.9

31

4.3

30

0.6

10000         

16

3.1

28

1.2

25

2.3

Positive Control

1320

21.1

864

24.3

697

11.5

 

Strain TA1537 

Dose

No Activation
(Negative)

10% HLI
(Negative)

10% RLI
(Negative)

Dose units

Mean

± SEM

Mean

± SEM

Mean

± SEM

0         

8

0.3

8

0.7

6

2.3

100         

4

0.6

7

1

10

2.9

333         

6

0.6

11

3.2

9

0.6

1000         

8

0.9

11

1.8

9

2

3333         

6

1.3

10

1.5

8

1.7

10000         

7

0.3

11

0.6

10

0.9

Positive Control

282

11.2

88

7

82

6

 

Abbreviations:
RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9

Applicant's summary and conclusion