Hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Remarks:

the purity and composition of the substance tested are not characterized, therefore it can not be excluded that the effects are caused by an impurity

Data source

Reference

Reference Type:

publication

Title:

Chlorotriazine Reactive Azo Red 120 Textile Dye Induces Micronuclei in Fish

Author:

Kabil Al-Sabti

Year:

2000

Bibliographic source:

Ecotoxicology and Environmental Safety 47, 149}155 (2000)

Report date:

1999

Materials and methods

Test guideline

Qualifier:

no guideline available

GLP compliance:

no

Type of assay:

other: in vivo micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

Hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

EC Number:

269-284-3

EC Name:

Hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

Cas Number:

68214-04-0

Molecular formula:

C44H24Cl2N14Na6O20S6

IUPAC Name:

hexasodium 4,4'-[1,4-phenylenebis[imino(6-chloro-1,3,5-triazine-4,2-diyl)imino]]bis[5-hydroxy-6-[(2-sulphonatophenyl)azo]naphthalene-2,7-disulphonate]

Details on test material:

no details provided (structure depicted in the publication)

Test animals

Species:

other: fish

Sex:

not specified

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Species: Prussian carp, Carassius auratus gibelio,
- Source: Ribe Maribor Fish farm, in Race near Maribor in Slovenia
- Length at study initiation: 12 cm
- Weight at study initiation: 100 g
- Feeding: witheld 2 days before start of treatment, fed during the test
- Water: fresh water
- Acclimation period: 7 days at a population density of 100 specimens in a 400 L tank with well-aerated nonchlorinated water at 15°C and pH 7

ENVIRONMENTAL CONDITIONS: no data

Administration / exposure

Vehicle:

none

Duration of treatment / exposure:

exposure for 9 days

Frequency of treatment:

continuously (concentrations were assessed photometrically and the concentration was adjusted when necessary to maintain it constant)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

1 other: mg/L nominal

Doses / concentrations 2

Dose / conc.:

5 other: mg/L nominal

Doses / concentrations 3

Dose / conc.:

10 other: mg/L nominal

No. of animals per sex per dose:

7 fish/concentration

Control animals:

yes

Positive control(s):

10 ppm benzene

Examinations

Tissues and cell types examined:

erythrocytes

Details of tissue and slide preparation:

Before blood sampling, fish were tagged by cutting one or a combination of two fins to enable blood to be sampledfrom the same fish specimen at 3, 6, and 9 days (time response) while exposed to the same concentration of CRARD-120 and from both negative and positive control groups. Blood samples for the smears were obtained by caudal vein puncture of the same fish specimen at three intervals, and slides were prepared by the fish micronucleated erythrocyte method (Al-Sabti, 1986). From each fish, two slides were prepared. After fixation in pure ethanol for 20 min, the prepared slides were left to air dry, and then the smears were stained with 5% Giemsa solution for 20 min. The slides were examined by light microscopy (Opton, Germany) under an oil immersion lens. Scoring of coded slides was done "blind.'' Only erythrocyte cells that were
isolated from surrounding cells were scored, and MNs were enumerated only if they were distinct from the two nuclei. One thousand erythrocytes per specimen for each slide (2000 per two slides) were analyzed in the subsequent treatments to determine the frequency of cells with one or two micronuclei.

Statistics:

method of Becker et al. (mean and SD)

Results and discussion

Test results

Key result

Sex:

not specified

Genotoxicity:

positive

Toxicity:

no effects

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Micronuclei were elevated not only in a dose-dependent (1, 5, and 10 mg/L) but also in a time-dependent (3, 6, and 9 days) manner, compared with the negative (tap water) and positive (10 ppm benzene) control groups.

Any other information on results incl. tables

Test group	day	Number of fish	Micronuclei per 1000 Fish Erythrocytes
control	3	5	4.7±0.6
benzene	3	7	8.36±0.35
1 mg/L	3	6	9.8±2.1
5 mg/L	3	7	16.1±2.1
10 mg/L	3	6	19.3±1.0
control	6	5	5.2±0.3
benzene	6	7	15.07±0.56
1 mg/L	6	6	12.3±1.1
5 mg/L	6	7	18.9±1.4
10 mg/L	6	6	22.8±0.8
control	9	5	6.2±0.4
benzene	9	7	24.2±0.7
1 mg/L	9	6	15.5±1.2
5 mg/L	9	7	20.6±1.7
10 mg/L	9	6	24.6±1.4

 

Applicant's summary and conclusion

Conclusions:

Under the conditions of the test the substance induced micronuclei in erythrocytes of fish

Executive summary:

Micronucleus induction in fish erythrocytes was used to study the risk to aquatic ecosystems due to the genotoxicity of Chlorotriazine Reactive Azo Red 120 textile dye. The frequencies of micronuclei were studied for three low doses of 1, 5, and 10 mg/L and blood sampling was carried out on the same 5sh after 3, 6, and 9 days. It was found that micronuclei increased not only in a dose-dependent manner but also in a time-dependent way, compared with negative (tap water) and positive (10 ppm benzene) control groups. There was also a slight, time-dependent increase in erythrocyte micronuclei of the control fish specimens.