.beta.-Alanine, N-[2-chloro-4-[2-(4-nitrophenyl)diazenyl]phenyl]-, 2-[2-(1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl)ethoxy]-2-oxoethyl ester

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

From January 13 to February 19, 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Yellow 981

IUPAC Name:

Yellow 981

Test material form:

solid: particulate/powder

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Test concentrations with justification for top dose:

- Test concentrations, experiment I: 33, 100, 333, 1000, 2500 and 5000 µg/plate
- Test concentrations, experiment II: 10, 33, 100, 333, 1000 and 2500 µg/plate
- Justification: pre-experiment for toxicity

Vehicle / solvent:

DMSO

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

methylmethanesulfonate

other: 4-nitro-o-phenylene-diamine: TA 1537 (50 µg/plate) and TA 98 (10 µg/plate)

Remarks:

without metabolic activation

Controls 2

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene: TA 1535 (2.5 µg/plate), TA 1537 (2.5 µg/plate), TA 98 (2.5 µg/plate), TA 100 (2.5 µg/plate), TA 102 (10 µg/plate)

Remarks:

with metabolic activation

Details on test system and experimental conditions:

Plate incorporation test

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

in exp. I and II

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

in exp. I with S9 and in exp. II without S9

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

in exp. I and II; both with and without S9

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

in exp. I with S9 only

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

in exp. I and II

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

The plates incubated with the test item showed normal background growth up to the highest concentration with and without metabolic activation in both independent experiments.
Toxic effects, evident as a reduction in the number of revertants, occurred at the higher concentrations in nearly all strains without S9 mix in both experiments. ln the presence of metabolic activation toxic effects were observed in strain TA 102 in experiment I and in strain TA 1535 in experiment ll.
ln experiment l, substantial and dose dependent increases in revertant colony numbers were observed following treatment with test item in strains TA 1537, TA 98, and TA 100 with metabolic activation and in strain TA 98 without metabolic activation. ln experiment ll a dose dependent increase in revertant colony numbers was observed without S9 mix in strains TA 1537 and TA 98 and with S9 mix in strain TA 98.
Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.

Applicant's summary and conclusion

Conclusions:

Positive: induces gene mutations by base pair changes and frameshifts in strains TA 1537, TA 98, and TA 100 in the bacterial reverse mutation assay.