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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: data from peer - reviewed journals

Data source

Reference
Reference Type:
publication
Title:
Genotoxicity Testing of the Food Colours Amaranth and Tartrazine
Author:
Aparajita Das and Anita Mukherjee
Year:
2004
Bibliographic source:
Int J Hum Genet, 4(4): 277-280 (2004)

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)
Principles of method if other than guideline:
Chromosome aberration assay was performed to evaluate the mutagenic nature of the test compound Amaranth.
GLP compliance:
not specified
Type of assay:
endogenous gene animal assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate
EC Number:
213-022-2
EC Name:
Trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate
Cas Number:
915-67-3
Molecular formula:
C20H14N2O10S3.3Na
IUPAC Name:
trisodium 3-hydroxy-4-(4'-sulphonatonaphthylazo)naphthalene-2,7-disulphonate
Constituent 2
Reference substance name:
trisodium (4E)-3-oxo-4-[(4- sulfonato-1- naphthyl)hydrazono]naphthalene- 2,7-disulfonate
IUPAC Name:
trisodium (4E)-3-oxo-4-[(4- sulfonato-1- naphthyl)hydrazono]naphthalene- 2,7-disulfonate
Constituent 3
Reference substance name:
Amaranth dye
IUPAC Name:
Amaranth dye
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Name of test material (as cited in study report): Amaranth dye
Molecular formula (if other than submission substance): C20H11N2Na3O10S3
Molecular weight (if other than submission substance): 604.47
Smiles notation (if other than submission substance): c1ccc2c(c1)c(ccc2S(=O)(=O)[O])N=Nc3c4ccc(cc4cc(c3O)S(=O)(=O)[O-])S(=O)(=O)[O-].[Na+].[Na+].[Na+]
InChl (if other than submission substance): 1S/C20H14N2O10S3.3Na/c23-20-18(35(30,31)32)10-11-9-12(33(24,25)26)5-6-13(11)19(20)22-21-16-7-8-17(34(27,28)29)15-4-2-1-3-14(15)16;;;/h1-10,23H,(H,24,25,26)(H,27,28,29)(H,30,31,32);;;/q;3*+1/p-3
Substance type: Organic
Physical state: Solid

Test animals

Species:
mouse
Strain:
other: Swiss Albino
Sex:
male
Details on test animals or test system and environmental conditions:
Details on test animals and env conditions
TEST ANIMALS
Source: No data available
Age at study initiation: 8-10 weeks
Weight at study initiation: 20-25 g
Assigned to test groups randomly: [no/yes, under following basis: ] No data available Fasting period before study: No data available
Housing: No data available
Diet (e.g. ad libitum): Commercial diet, ad libitum
Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
Temperature (°C): Ambient room temperature
Humidity (%): Relative
Air changes (per hr): No data available
Photoperiod (hrs dark / hrs light): No data available

IN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
Vehicles
Vehicle(s)/solvent(s) used: distilled water
Justification for choice of solvent/vehicle: No data available
Concentration of test material in vehicle: 50,100 and 200 mg/kg body weight
Duration of treatment / exposure:
18 hours
Doses / concentrations
Remarks:
Doses / Concentrations:
50,100 and 200 mg/kg bw
Basis:
no data
No. of animals per sex per dose:
Total : 16
0 mg/kg bw: 4 male mice
50 mg/kg bw: 4 male mice
100 mg/kg bw: 4 male mice
200 mg/kg bw: 4 male mice
Control animals:
yes, concurrent vehicle
Positive control(s):
Positive controls : cyclophosphamide CP
Doses / concentrations: 20 mg/Kg bw

Examinations

Tissues and cell types examined:
Bone marrow cells
Details of tissue and slide preparation:
Details of tissue and slide preparation
CRITERIA FOR DOSE SELECTION: The selection of dose was based on the permitted dose of the dyes being 100mg/kg of the prepared food.
TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):

DETAILS OF SLIDE PREPARATION: Bone marrow cells were routinely processed by the standard procedure and slides were coded and stained in diluted Giemsa

METHOD OF ANALYSIS: For chromosomal aberration analysis, four animals were used per point. Hundred well spread metaphase plates were scored per animal (400 metaphase plates per treatment set) at random.

OTHER: All aberrations (chromatid gaps, isochromosome gaps, chromatid breaks and rearrangements) were considered equal- regardless of the number of breakages involved. The percentages of damaged cells (% DC) and chromosomal aberrations per cell (CA/cell) values were calculated excluding gaps.
Evaluation criteria:
Chromatid gaps, isochromosome gaps, chromatid breaks and rearrangements were noted
Statistics:
ANOVA test was performed at 0.05 level

Results and discussion

Test results
Sex:
male
Genotoxicity:
negative
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
valid

Any other information on results incl. tables

Table 1: Chromosomal aberrations of mice bone marrow cells following treatment with different doses of Amaranth

 

Dose

mg/kg

Type of Aberrations

%DC

(MEAN±SEM)

 

 

%CA/Cell

(MEAN±SEM)

G’

G’’

 

B’

 

B’’

 

RR

 

50

7

1

2

-

-

2.00± 1.00

0.02± 0.01

100

10

1

5

-

-

3.50± 0.50

0.03± 0.00

200

8

2

6

2

-

4.00± 0.81

0.04± 0.01

 

Where: G’ G’’ = Chromatid and chromosome gaps

B’ B’’ = Chromatid and chromosome breaks

RR = rearrangements,

DC = Percentage of damaged cells

CA/Cell = Chromosome aberration

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
The test compound amaranth is not mutagenic in the chromosomal aberration study conducted using Swiss albino mouse isolated bone marrow cells.
Executive summary:

Chromosome aberration assay was performed to evaluate the mutagenic nature of the test compound amaranth.

 

The study was conducted on male Swiss albino mice, 8-10 weeks old and weighing 20-25 g. Four animals per dose were administered intraperitoneally with the different doses of the amaranth (50,100 and 200 mg/kg body weight).

The animals were killed after 18 hr. For bone marrow chromosome analysis, animals were injected with 0.1 ml colchicine solution (4mg/10ml distilled water /10g body weight, 90 minutes before they were killed. Bone marrow cells were routinely processed by the standard procedure and slides were coded and stained in diluted Giemsa.

 

For chromosomal aberration analysis, four animals were used per point. Hundred well spread metaphase plates were scored per animal (400 metaphase plates per treatment set) at random. The types of aberrations were scored. The percentages of damaged cells (% DC) and chromosomal aberrations per cell (CA/cell) values were calculated excluding gaps.

 

The test compound amaranth is not mutagenic in the chromosomal aberration study conducted using Swiss albino mouse isolated bone marrow cells.

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