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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
In vitro study of DNA damage induced by acid orange 52 and its biodegradation derivatives
Author:
HEDI BEN MANSOUR,†‡ DANIEL BARILLIER,† DAVID CORROLER,† KAMEL GHEDIRA,‡ LEILA CHEKIR-GHEDIRA, *‡§ and RIDHA MOSRATI†
Year:
2009
Bibliographic source:
Environmental Toxicology and Chemistry, Vol. 28, No. 3, pp. 489–495, 2009

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
In vitro gene toxicity study of acid orange 52 (AO52) in Salmonella Typhimurium TA102 and TA104
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Acid orange 52 (AO52) - Molecular formula (if other than submission substance): C14-H15-N3-O3-S.Na- Molecular weight (if other than submission substance): 327.3386 g/mole - Substance type: Organic - Physical state: No data available Purity: No data available - Impurities (identity and concentrations): No data available

Method

Target gene:
No data
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA102 and TA104
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254
Test concentrations with justification for top dose:
250, 50, and 10 g/plate
Vehicle / solvent:
No data available
Controls
Untreated negative controls:
yes
Remarks:
spontaneous revertants.
Negative solvent / vehicle controls:
not specified
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: With S9 - 5 g/plate of 2-AA Without S9 - 86.5 μg/plate of methyl methane Sulfonate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)DURATION- Preincubation period: 45 min- Exposure duration: 48 h- Expression time (cells in growth medium): 16 h- Selection time (if incubation with a selection agent): No data available - Fixation time (start of exposure up to fixation or harvest of cells): No data available SELECTION AGENT (mutation assays): No data available SPINDLE INHIBITOR (cytogenetic assays): No data available STAIN (for cytogenetic assays): No data available NUMBER OF REPLICATIONS: No data available NUMBER OF CELLS EVALUATED: 2 DETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No data available OTHER EXAMINATIONS:- Determination of polyploidy: No data available - Determination of endoreplication: No data available - Other: No data available OTHER: An overnight culture of bacteria (100 μl, cultivated for 16 h at 37°C, approximate cell density 2 X 108 cells/ml) and sodium phosphate buffer (500 μl, 0.2 M, pH 7.4, for assay without metabolic activation system) or S9 mix (500 μl) were distributed in sterilized capped tubes in an ice bath, then 100 μl of test concentration 250, 50, and 10 g/plate was added.
Evaluation criteria:
Revertant bacterial colonies were examined.
Statistics:
Mutagenic effect data are expressed as mean ± standard deviation from three replicates. The statistical analyses were performed with Statistica ’99 Edition (Stat Soft France, Maisons- Alfort, France). The Duncan test was used to compare test compounds with the negative control (spontaneous revertants), and the difference was considered to be significant at p < 0.05.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium, other: TA102
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: TA104
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
No data
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Mutagenic activity of AO52 (after static or shaken degradation with Pseudomonas putida mt-2), 4-ABS, and DMPD evaluated by the Salmonella Typhimurium TA104 and TA102 assay systems with and without the metabolic activation system (S9)

 

 

Mutagenic activity (revertants/plates)b

 

 

TA104

TA102

Tested compounda

Dose

(μg/plate)

-S9

+S9

               -S9

+S9

PC

1,846 ± 46***

1,400 ± 53***

1,721 ± 24***

1,031 ± 231***

SP

203 ± 11

273 ± 14

175 ± 10

217 ± 14

Pure AO52

250

221 ± 09

755 ± 24**

254 ± 09

639 ± 21**

 

50

198 ± 08

555 ± 25

222 ± 08

544 ± 19

 

10

197 ± 04

382 ± 23

213 ± 11

311 ± 14

a4-ABS = 4-aminobenzenesulfonic acid; AO52 = acid orange 52; DMPD = N,N’-dimethyl-p-phenylenediamine; PC = positive control; PPD = p-phenylenediamine; SP = spontaneous revertants.

bTA102/–S9 and TA104/–S9 = methyl methane sulfonate (86.5 μg/plate); TA102/+S9 and TA104/+S9 =2-aminoanthracene (5 μg/plate).

Values are significant at * p < 0.01, ** p < 0.001, and *** p < 0.0001.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):negative without metabolic activationpositive with metabolic activationAcid orange 52 (AO52) was considered to be negative without S9 and positive gene toxic with S9 metabolic activation when Salmonella Typhimurium TA102 and TA104 tested.
Executive summary:

In a in vitro gene mutation test, Salmonella Typhimurium TA102 and TA104 was tested for Revertant bacterial colonies by using Acid orange 52 (AO52) with and without S9 activation in the concentration of 250, 50 and 10 g/plate. Revertant bacterial colonies were not observed in without S9 activation and with S9 metabolic activation revertant bacterial colonies were observed. Therefore, Acid orange 52 (AO52) was considered to be negative without S9 and positive gene toxic with S9 metabolic activation when Salmonella Typhimurium TA102 and TA104 were tested.