Registration Dossier

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Remarks:
No reference to OECD guideline. Deviation from OECD guideline with the use of 4-o-Tolylazo-o-toluidine as positive control. Study conducted in 1975.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1975
Report date:
1975

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
Only 4 strains of bacteria were used. Strain TA1538 was used and Strain TA98 was not. Positive controls deviate from those recommended in guideline.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
m-phenylenediamine
EC Number:
203-584-7
EC Name:
m-phenylenediamine
Cas Number:
108-45-2
Molecular formula:
C6H8N2
IUPAC Name:
m-phenylenediamine
Details on test material:
-Purity: not reported
-Name of test material (as cited in study report): m-phenylenediamine

Method

Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535
Species / strain / cell type:
S. typhimurium TA 1537
Species / strain / cell type:
S. typhimurium TA 1538
Species / strain / cell type:
S. typhimurium TA 100
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbitol-stimulated rat liver homogenate
Test concentrations with justification for top dose:
1, 10, 50, 100, 250, 500, 750, 1000 micrograms/plate.
Vehicle / solvent:
no data
Controls
Untreated negative controls:
yes
Positive controls:
yes
Positive control substance:
other: 4-o-Tolylazo-o-toluidine (for TA1538 and metabolic activation) and N-methyl-N'-nitro-N-nitrosoguanidine
Details on test system and experimental conditions:
In the absence of metabolic activation, approximately 5E8 bacteria were added to 2 mL of top agar (0.6% Difco agar, 0.6% NaCl, 0.050 mM of L-histidine, 0.05 mM of biotin). After the test chemical was added to the top agar, the solution was mixed and poured onto the surface of a minimal agar plate with Vogel-Bonner E medium containing 1.5% agar and 2% glucose.

The metabolic activation system consisted of 0.15 mL of the 9000 x g supernatant of homogenized rat liver, MgCl2, KCl, glucose-6-phosphate, TPN, and a sodium phosphate buffer (pH 7.4). This mixture (0.25 mL) was added directly to the top agar immediately before it was poured over the minimal agar plate. The plates were incubated at 37 deg C for 40 hours. The number of histidine-positive (his+) revertant colonies on each plate then was counted. All tests were performed in duplicate.

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION: 40 hours

SELECTION AGENT (mutation assays): histidine

NUMBER OF CELLS EVALUATED: 5E8

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified

Any other information on results incl. tables

In Vitro Salmonella Typhimurium Assay With m-PDA.

 

Chemical

Metabolic Activation

Amount of Compound Added per Plate

(µg)

Average Number of Histidine-Positive Revertants per Plate

TA100

TA1535

TA1537

TA1538

Negative Control

-

 

72

11

5

9

+

 

84

11

6

15

 

 

 

 

 

 

 

4-o-Tolylazo-o-toluidine

(Positive control for TA1538 and metabolic activation)

-

25

 

 

 

10

+

25

 

 

 

195

 

 

 

 

 

 

 

N-methyl-N’-nitro-N-nitrosoguanidine

-

2

 

600

 

 

 

 

 

 

 

 

 

m-phenylenediamine

-

1

65

9

4

4

 

-

10

64

8

4

2

 

-

50

78

9

5

9

 

-

100

80

10

3

12

 

-

250

 

 

 

9

 

-

500

83

8

2

17

 

-

750

 

 

 

 

 

-

1000

74

11

5

21

 

+

1

87

14

8

10

 

+

10

78

11

7

17

 

+

50

80

9

6

79

 

+

100

85

14

6

177

 

+

250

 

 

 

423

 

+

500

84

14

11

570

 

+

750

 

 

 

623

 

+

1000

79

10

8

971

 

Applicant's summary and conclusion

Conclusions:
The test substance was mutagenic in strain TA 1538 with metabolic activation in the S. typhimurium assay, but negative when activation was not used. It was negative in all other strains, with or without activation.
Executive summary:

The test substance was mutagenic in strain TA 1538 with metabolic activation in the S. typhimurium assay, but negative when activation was not used. It was negative in all other strains, with or without activation.

Categories Display