2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

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:

2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study performed according to OECD guideline 471 in compliance to GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

N/A

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

EC Number:

251-410-3

EC Name:

2,2'-[[4-[(2-hydroxyethyl)amino]-3-nitrophenyl]imino]bisethanol

Cas Number:

33229-34-4

Molecular formula:

C12H19N3O5

IUPAC Name:

2-({4-[bis(2-hydroxyethyl)amino]-2-nitrophenyl}amino)ethan-1-ol

Details on test material:

Test item : HC Blue No 2
EC number : 251-410-3
Batch number : 118pt 2

Method

Target gene:

Histidine gene

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

other: see below

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Details on mammalian cell type (if applicable):

N/A

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9

Test concentrations with justification for top dose:

Experiment 1 :
50, 150, 500, 1500 and 5000 ug/plate

Experiment 2 :
50, 150, 500, 1500 and 5000 ug/plate for all of the tester strains except TA98 which was allocated an amended dose range of 150, 500, 1500, 3000 and 5000 ug/plate. The intermediate dose level (3000 ug/plate) was included to achieve a better dose-response relationship.

Vehicle / solvent:

Solvent (DMSO)

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

TA100 and TA1535 (both without S9)

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 (without S9)

Controls 3

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

mitomycin C

Remarks:

TA102 (without S9)

Controls 4

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

TA98 (without S9)

Controls 5

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

TA100, TA1535, TA1537 (all with S9)

Controls 6

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Remarks:

TA98 (with S9)

Controls 7

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 1,8dihydroxyanthraquinone

Remarks:

TA102 (with S9)

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium, other: TA1535, TA 1537, TA102, TA100

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:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

positive

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA102, TA100

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

Remarks on result:

other: other: experiment 1

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive without metabolic activation

Under the experimental conditions used in this study, HC Blue No 2 was considered to be genotoxic (mutagenic) in Salmonella typhimurium strain TA98 in the absence of metabolic activation.

Executive summary:

Salmonella typhimurium strains TA1535, TA1537, TA102, TA98 and TA100 were treated with the test material HC Blue No 2 using the Ames plate incorporation method method at five dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system. The dose range was determined in a premiminary toxicity assay and was 50 to 5000 ug/plate in the first experiment. The experiment was repeated on a separate day using the same dose range as experiment 1 for all tester strains except TA98, fresh cultures of the bacterial strains and fresh test material formulations. The dose range for tester strain TA98 was amended slightly following information obtained from experiment 1 and was 150, 500, 1500, 3000 and 5000 ug/plate. The intermediate dose level of 3000 ug/plate was included in an attempt to establish a better dose-response relationship. The vehicle control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without metabolic activation. The sensitivity of the assay and the efficacy of the S9 mix were validated. Statistically significant, dose related and reproducible increases in the frequency of revertant colonies were observed in tester strain TA98, without metabolic activation, at the upper dose levels of the test material. No significant increases in the frequency of revertant colonies were recorded for any of the remaining bacterial strains, with any dose of the test material, either with or without metabolic activation. Under the experimental conditions used in this study, HC Blue No 2 was considered to be genotoxic (mutagenic) in Salmonella typhimurium strain TA98 in the absence of metabolic activation.