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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1987
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Referenceopen allclose all

Reference Type:
publication
Title:
Chromosome aberrations and sister chromatid exchanges in chinese hamster ovary cells: evaluation of 108 chemicals.
Author:
Galloway SM, Armstrong MJ, Reuben C, Colman S, Brown B,|Cannon C, Bloom AD, Nakamura F, Ahmed M, Duk S, Rimpo J,|Margolin BH, Resnick MA, Anderson B, Zeiger E
Year:
1987
Bibliographic source:
Environ Mol Mutagen 10 [Suppl.10]: 1-175 (1987)
Report date:
1987
Reference Type:
publication
Title:
Unnamed
Year:
1993

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Deviations:
yes
GLP compliance:
not specified
Type of assay:
in vitro mammalian chromosome aberration test

Test material

Constituent 1
Chemical structure
Reference substance name:
1-chloro-4-nitrobenzene
EC Number:
202-809-6
EC Name:
1-chloro-4-nitrobenzene
Cas Number:
100-00-5
Molecular formula:
C6H4ClNO2
IUPAC Name:
1-chloro-4-nitrobenzene
Details on test material:
- Name of test material (as cited in study report): 1-chloro-4-nitrobenzene
- Analytical purity: > 99 %
Specific details on test material used for the study:
- Name of test material (as cited in study report): 1-chloro-4-nitrobenzene
- Analytical purity: > 99 %

Method

Target gene:
Not applicable
Species / strain
Species / strain / cell type:
other: Chinese hamster ovary cells
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9-Mix
Test concentrations with justification for top dose:
- S9: (1) 0, 50, 167, 500 µg/ml, harvest time (ht): 10.5 hrs; (2) 0, 700, 800, 900 µg/ml, ht: 10.6 hrs; (3)0, 500, 600, 700 µg/ml, ht: 19.0 hrs;
+ S9: (4) 0, 50, 167, 500, 5000 µg/ml, ht: 10.5 hrs; (5) 0, 600, 800, 900 µg/ml, ht: 19 hrs
Vehicle / solvent:
DMSO
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
mitomycin C
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
cyclophosphamide
Statistics:
statistical procedures according to Galloway et al. (1985) and Margolin et al. (1986)

Results and discussion

Test results
Species / strain:
Chinese hamster Ovary (CHO)
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
- dose selection based on preliminary growth inhibition test, or based on observations of cell confluence and mitotic cell availability in the SCE test, performed in the same laboratory
- The aberration test without activation gave a high frequency of breaks in two of the three test and at an extremely toxic dose. The evaluation was based on only 33 and 22 metaphases. Positive results were obtained in the second of the two tests with activation by using closely spaced, high doses (19 hours harvest time). Cells with aberrations were up to 31% increased compared to solvent control were observed.

Any other information on results incl. tables

Tabel: Induction of chromosomal aberrations in CHO by 4-chloronitrobenzene

 

-S9

 

+S9

 

Dose (µ/ml)

Total Cells

No. of Abs

Abs/Cell

Cells with Abs (%)

 

Dose (µ/ml)

Total Cells

No. of Abs

Abs/Cell

Cells with Abs (%)

Trial 1 - Harvest time: 10,5h

Summary: Negative

 

DMSO

 

 

Mitomycin-C

 

4-Chloronitrobenzene

 

 

 

 

 

 

 

1

 

50

167

500

 

 

 

 

100

100

 

100

 

100

100

100

 

 

 

 

2

1

 

22

 

1

2

0

 

 

 

 

0,02

0,01

 

0,22

 

0,01

0,02

0,00

 

 

 

 

2,0

1,0

 

19,0

 

1,0

2,0

0,0

 

P=0,665

Trial 1 - Harvest time: 10,5h

Summary: Negative

 

 

DMSO

 

Cyclophosphamide

 

4-Chloronitrobenzene

 

 

 

 

 

 

 

25

 

50

167

500

5000

 

 

 

 

 

100

 

100

 

100

100

100

100

 

 

 

 

 

4

 

24

 

4

5

1

2

 

 

 

 

 

0,04

 

0,24

 

0,04

0,05

0,01

0,02

 

 

 

 

 

4,0

 

20,0

 

4,0

5,0

1,0

2,0

 

P=0,881

Trial 2 - Harvest time: 10,6h

Summary: Weakly Positive

 

DMSO

 

 

Mitomycin-C

 

4-Chloronitrobenzene

 

 

 

 

 

 

 

 

1

 

700

800

900

 

 

 

 

 

100

100

 

100

 

100

100

100

 

 

 

 

 

2

4

 

16

 

1

5

8

 

 

 

 

 

0,02

0,04

 

0,16

 

0,01

0,06

0,24

 

 

 

 

 

2,0

3,0

 

16,0

 

1,0

6,0

24,0

 

P<0,001

Trial 2 - Harvest time: 19h

Summary: Positive

 

 

DMSO

 

Cyclophosphamide

 

4-Chloronitrobenzene

 

 

 

 

 

 

 

50

 

600

800

900

 

 

 

 

 

100

 

50

 

35

100

100

 

 

 

 

 

0

 

236

 

6

38

4

 

 

 

 

 

0,00

 

4,72

 

0,17

0,38

0,04

 

 

 

 

 

0,0

 

88,0

 

17,0

31,0

3,0

 

 

P=0,002

Trial 3 - Harvest time: 19h

Summary: Weakly Positive

 

DMSO

 

 

Mitomycon-C

 

4-Chloronitrobenzene

 

 

 

 

 

 

 

 

1

 

500

600

700

 

 

 

 

 

100

100

 

100

 

100

50

22

 

 

 

 

 

1

0

 

26

 

0

0

1

 

 

 

 

 

0,01

0,00

 

0,26

 

0,00

0,00

0,05

 

 

 

 

 

1,0

0,0

 

21,0

 

0,0

0,0

5,0

 

P=0,013

 

 

 

 

 

 


(1) neg. (2) weak pos. (3) weak pos.
(4) neg. (5) pos.
The aberration test without activation gave a high frequency
of breaks in two of the three tests and at an extremely
toxic dose. The high point was based on only 33 and 22
metaphases.
Positive results were obtained in the second of the two
tests with activation by using closely spaced, high doses.
Cells with aberrations up to 31% increased compared to
solvent control

Applicant's summary and conclusion

Conclusions:
Interpretation of results : positive
In conclusion, the test item is considered to be clastogenic in this chromosome aberration test.
Executive summary:

Galloway et al., 1987

p-Chloronitrobenzol was tested in vitro to determine whether it would cause chromosomal aberrations in a mammalian cell line derived form Chinese hamster ovary tissue according to OECD guideline 473. The cells were incubated with different concentrations. With S9 -mix with 0, 50, 167, 500, 5000 µg/ml, ht: 10.5 hrs; (5) 0, 600, 800, 900 µg/ml, ht: 19 hrs and without S9-mix with 0, 50, 167, 500 µg/ml, harvest time (ht): 10.5 hrs; (2) 0, 700, 800, 900 µg/ml, ht: 10.6 hrs; (3) 0, 500, 600, 700 µg/ml, ht: 19.0 hrs.

The aberration test without activation gave a high frequency of breaks in two of the three test and at an extremely toxic dose. The evaluation was based on only 33 and 22 metaphases. Positive results were obtained in the second of the two tests with activation by using closely spaced, high doses (19 hours harvest time). Cells with aberrations were up to 31% increased compared to solvent control were observed.