1-chloro-4-nitrobenzene

Genetic toxicity: in vitro

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

Reference

Reference 1

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 2

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

Test material information

Constituent 1

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

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

mitomycin C

Controls 2

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.