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Toxicological information

Genetic toxicity: in vivo

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Administrative data

Endpoint:
genetic toxicity in vivo
Remarks:
Type of genotoxicity: genome mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Repeated-dose liver and gastrointestinal tract micronucleus assays with CI Solvent Yellow 14 (Sudan I) using young adult rats
Author:
Shoji Matsumura, Naohiro Ikeda, Shuichi Hamada, Wakako Ohyama, Yumi Wako, Kazufumi Kawasako, Toshio Kasamatsu, Naohiro Nishiyama
Year:
2015
Bibliographic source:
Mutation Research 780-781 (2015) 76–80

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
Bone marrow MN micronucleus (MN) assay was performed to determine the mutagenic nature of Solvent yellow 14
GLP compliance:
not specified
Type of assay:
other: Bone marrow MN micronucleus (MN) assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1-phenylazo-2-naphthol
EC Number:
212-668-2
EC Name:
1-phenylazo-2-naphthol
Cas Number:
842-07-9
Molecular formula:
C16H12N2O
IUPAC Name:
1-[(E)-2-phenyldiazen-1-yl]naphthalen-2-ol
Test material form:
solid: crystalline
Details on test material:
- Name of test material (as cited in study report): Solvent yellow 14
- Molecular formula (if other than submission substance): C16H12N2O
- Molecular weight (if other than submission substance): 248.28
- InChl (if other than submission substance):1S/C16H12N2O/c19-15-11-10-12-6-4-5-9-14(12)16(15)18-17-13-7-2-1-3-8-13/h1-11,19H/b18-17+
- Substance type: Organic
- Physical state: Solid
- Analytical purity: >95%
- Impurities (identity and concentrations): No data
Specific details on test material used for the study:
- Name of test material: Solvent yellow 14
- Molecular formula: C16H12N2O
- Molecular weight: 248.284 g/mol
- Substance type: Organic
- Physical state: Solid
- Purity: >95%
- Impurities (identity and concentrations): No data available

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Details on species / strain selection:
No data
Sex:
male
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Japan Inc. (Yokohama,Japan)
- Age at study initiation: 6 weeks old
- Weight at study initiation: No data available
- Assigned to test groups randomly: No data available
- Fasting period before study: No data available
- Housing: The animals were housed in an air-conditioned room
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%): No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12 h light/dark cycle

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

Administration / exposure

Route of administration:
oral: unspecified
Vehicle:
- Vehicle(s)/solvent(s) used: yes- methyl cellulose
- Justification for choice of solvent/vehicle: No data available
- Concentration of test material in vehicle: 0, 150, 300, and 600 mg/kg/day
- Amount of vehicle (if gavage or dermal): No data available
- Type and concentration of dispersant aid (if powder): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: Sudan I was suspended in 0.5% (w/v) Methyl Cellulose Solution (Wako Pure Chemical Industries, Ltd., Osaka, Japan) each day before dosing to guve a dose range of 0, 150, 300 or 600 mg/Kg/day

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available
Duration of treatment / exposure:
14 days
Frequency of treatment:
Daily
Post exposure period:
No data available
Doses / concentrations
Remarks:
Doses / Concentrations:
0, 150, 300, and 600 mg/kg/day
No. of animals per sex per dose:
Total: 20
0 mg/Kg/day: 5 male rats
150 mg/Kg/day: 5 male rats
300 mg/Kg/day: 5 male rats
600 mg/Kg/day: 5 male rats
Control animals:
yes, concurrent vehicle
Positive control(s):
No data available

Examinations

Tissues and cell types examined:
Micronucleated Bone marrow cells
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION: Dose range finding study was conducted, The animals were dosed once daily for 7 days with 75, 150, 300, and 600 mg/kg/day of Sudan I and were monitored for clinical signs and body weight changes. A decrease in body weight was observed in the 600 mg/kg/day group compared with the vehicle control group, but no lethality or decrease in motor activity was observed. Based on these observations, the highest dose was set as 600 mg/kg/day and set the lower doses with the common ratio of two (300 and 150 mg/kg/day).

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): No data available

DETAILS OF SLIDE PREPARATION: smear preparations were made from the bone marroe cells and then stained with an AO solution and covered with a cover slip.

METHOD OF ANALYSIS: The specimens were observed under a fluorescent microscope with B-excitation (Blue light excitation), and the number of micronucleated immature erythrocytes (MNIMEs) per 2000 immature erythrocytes (IMEs) was counted for each animal.

OTHER: No data available
Evaluation criteria:
Micronucleated Bone marrow cells were observed
Statistics:
Differences in the incidences of MNHEPs between the test and the vehicle control groups were analyzed using the conditional binomial test reported by Kastenbaum and Bowman at the upper-tailed significance levels of 5% and 1%. The exception was that a Chi-squared test
was performed to analyze differences in the incidences of micronucleated immature erythrocytes (MNIMEs) between the test groups and the vehicle-treated control group.

The data were also statistically analyzed by a multiple comparison test according to the requirements of the organizing committee. Specifically, the homogeneity of the variance was examined using Bartlett’s test. When a homogeneous variance was demonstrated, one-way analysis of variance was applied. In cases of heterogeneous variance, the Kruskal–Wallis test was applied. When a statistically significant difference was demonstrated between groups, the difference was assessed by Dunnett’s test or a Dunnett-type multiple comparison test.

Results and discussion

Test results
Sex:
male
Genotoxicity:
positive
Toxicity:
not specified
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
not specified
Additional information on results:
RESULTS OF RANGE-FINDING STUDY
- Dose range: 75, 150, 300, and 600 mg/kg/day
- Solubility: No data available
- Clinical signs of toxicity in test animals: A decrease in body weight was observed in the 600 mg/Kg/bw dose level - Evidence of cytotoxicity in tissue analyzed: No data available
- Rationale for exposure: No data available
- Harvest times: No data available
- High dose with and without activation: No data available
- Other:No data available

RESULTS OF DEFINITIVE STUDY
- Types of structural aberrations for significant dose levels (for Cytogenetic or SCE assay): No data available
- Induction of micronuclei (for Micronucleus assay): No data available
- Ratio of PCE/NCE (for Micronucleus assay): No data available
- Appropriateness of dose levels and route: No data available
- Statistical evaluation: No data available

Any other information on results incl. tables

Results of the Bone marrow MN assay with Sudan:

 

Sudan I (mg/kg/day)

No. of animals

MNIMEs (%) individual

data (mean±SD)

0

5

0.60, 0.65, 0.75, 0.25, 0.50 (0.55±0.19)    

150

5

0.75, 1.25, 1.75, 1.10, 1.25 (1.22±0.35*)

300

5

0.65, 1.30, 2.00, 3.25, 1.65 (1.77±0.96*)

600

5

1.80, 2.40, 3.25, 1.75, 2.70 (2.38±0.63*)

* p < 0.01, Chi-squared test.

Applicant's summary and conclusion

Conclusions:
There was statistically significant increase in the incidences of MNIMEs observed with the repeated administration of 1-phenylazo-2-naphthol (Sudan I). Therefore in vivo genetic toxicity of 1-phenylazo-2-naphthol (Sudan I) is positive.
Executive summary:

The in vivo Geno toxicity of 1-phenylazo-2-naphthol (Sudan I) was examined using repeated-doseBone marrowmicronucleus (MN) assays in young adult maleCrl: CD (SD)rats.

They performed a dose range finding study. The animals were dosed once daily for 7 days with 75, 150, 300, and 600 mg/kg/day of Sudan I and were monitored for clinical signs and body weight changes. A decrease in body weight was observed in the 600 mg/kg/day group compared with the vehicle control group, but no lethality or decrease in motor activity was observed. Based on these observations, we determined the highest dose as 600 mg/kg/day and set the lower doses with the common ratio of two (300 and 150 mg/kg/day). BM cells were collected from the femurs. Immediately prior to microscopic observation, smear preparations were made with each sample and then stained with an AO solution and covered with a cover slip. The specimens were observed under a fluorescent microscope with B-excitation (Blue light excitation), and the number of micronucleated immature erythrocytes (MNIMEs) per 2000 immature erythrocytes (IMEs) was counted for each animal.

There was statistically significant increase in the incidences of MNIMEs observed with the repeated administration of Sudan I. Therefore in vivo genetic toxicity of 1-phenylazo-2-naphthol (Sudan I) is positive.

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