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Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
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:
Haematological Changes Induced By Feeding A Common Food Colour, Metanil Yellow, In Albino Mice
Author:
O.M. Prasad and Padam B. Rastogi
Year:
1983
Bibliographic source:
Toxicology Letters, 16 (1983) 103-107

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: Refer below principle
Principles of method if other than guideline:
Chronic oral toxicity study was performed to determine the oral toxic nature of metanil yellow upon repeated exposure
GLP compliance:
not specified
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
Sodium 3-(p-anilinophenylazo)benzenesulphonate
EC Number:
209-608-2
EC Name:
Sodium 3-(p-anilinophenylazo)benzenesulphonate
Cas Number:
587-98-4
Molecular formula:
C18H14N3O3S.Na
IUPAC Name:
sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
Test material form:
solid
Details on test material:
- Name of test material: Ext. D&C Yellow No. 1
- IUPAC name: sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
- Molecular formula: C18H14N3O3SNa
- Molecular weight: 375.383 g/mol
- Substance type: organic
Specific details on test material used for the study:
- Name of test material: Ext. D&C Yellow No. 1
- IUPAC name: sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
- Molecular formula: C18H15N3O3SNa
- Molecular weight: 375.383 g/mol
- Substance type: organic
- Physical state: No data
- Purity: No data available
- Impurities (identity and concentrations): no data available

Test animals

Species:
mouse
Strain:
other: Albino
Details on species / strain selection:
No data
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: No data
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 20-25 g
- Fasting period before study: No data
- Housing: No data
- Diet (e.g. ad libitum): pelleted lab chow (Hindustan Lever Ltd., Bombay, India)
- Water (e.g. ad libitum): Water ad libitum
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24°C ±1°C
- Humidity (%):No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data

IN-LIFE DATES: From: To: No data

Administration / exposure

Route of administration:
oral: feed
Details on route of administration:
No data
Vehicle:
other: Feed
Remarks:
pelleted lab chow (Hindustan Lever Ltd., Bombay, India)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Metanil yellow was mixed with pelleted lab chow (Hindustan Lever Ltd., Bombay, India) at dose levels of 0, 100, 500 or 3000 mg/Kg bw

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

VEHICLE
- Justification for use and choice of vehicle (if other than water): pelleted lab chow (Hindustan Lever Ltd., Bombay, India)
- Concentration in vehicle: 0, 100, 500 or 3000 mg/Kg bw
- Amount of vehicle (if gavage): No data
- Lot/batch no. (if required): No data
- Purity: No data
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
180 days
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
0, 100, 500 or 3000 mg/Kg bw
No. of animals per sex per dose:
Total: 320
0 mg/Kg bw: 40 males and 40 females
100 mg/Kg bw: 40 males and 40 females
500 mg/Kg bw: 40 males and 40 females
3000 mg/Kg bw: 40 males and 40 females
Control animals:
yes, concurrent vehicle
Details on study design:
No data
Positive control:
No data

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Throughout the experiment
- Cage side observations checked in table [No.?] were included. General condition of the animals was observed

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule: No data

BODY WEIGHT: No data
- Time schedule for examinations: No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: After 180 days
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. Total erythrocyte count (TEC), Differential leukocyte count (DLC)- neutrophils, lymphocytes, monocytes, eosinophils, basophils, hemoglobin, erythrocyte sedimentation rate (ESR), mean corpuscular volume (MCV), MCG, MCHC, Heinz bodies, platelet count (PC), PCV, Bleeding time (BT), coagulation time (CT)

CLINICAL CHEMISTRY: No data
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: No data
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data

OTHER: No data
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, organs were removed for gross pathology

HISTOPATHOLOGY: No data
Other examinations:
No data
Statistics:
No detailed data available

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
General condition was unaffected at dose levels of 100 and 500 mg/Kg bw except at 3000 mgKg bw
Mortality:
not specified
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
not specified
Ophthalmological findings:
not specified
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
100 mg/Kg bw: No abnormalilties were observed

500 mg/Kg bw: No abnormalilties were observed

3000 mg/Kg bw: Statistical analysis of various values exhibited significant decrease in TEC and Hb and increase in ESR, MCV and MCH suggesting normochromic macrocytic anaemia.

TLC, PC, PCV, MCHC, BT and CT, however, remained normal in both male and female animals.

A marked increase in the number of lymphocytes and macrocytes and a decrease in the number of neutrophils and eosinophils suggesting the occurrence of chronic lymphocytic leukaemia.

Heinz bodies began to appear in the blood after 3 months feeding in mice fed metanil yellow at 3.0 g/kg body weight and were observed in 80-90% erythrocytes at the end of the experiment.

Blood showed marked anisocytosis, poikilocytosis, stomatocytes, schistocytes and a large number of target cells, all normochromic in character. An
overwhelming preponderance of mature lymphocytes with scant cytoplasm, larger and denser clumps of nuclear chromatin and accentuation of parachromatin were also observed, which were suggestive of chronic lymphocytic leukaemia.
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Mice fed metanil yellow at 3000 mg/kg bw showed hepatomegaly and splenomegaly at autopsy.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Clinical signs and mortality
Clinical signs: General condition was unaffected at dose levels of 100 and 500 mg/Kg bw except at 3000 mgKg bw

Mortality: No data

Body weight and weight gain: No data

Food consumption and compound intake: No data

Food efficiency: No data

Water consumption and compound intake: No data

Opthalmoscopic examination: No data

Haematology:
100 mg/Kg bw: No abnormalilties were observed

500 mg/Kg bw: No abnormalilties were observed

3000 mg/Kg bw: Statistical analysis of various values exhibited significant decrease in TEC and Hb and increase in ESR, MCV and MCH suggesting normochromic macrocytic anaemia.

TLC, PC, PCV, MCHC, BT and CT, however, remained normal in both male and female animals.

A marked increase in the number of
lymphocytes and macrocytes and a decrease in the number of neutrophils and eosinophils suggesting the occurrence of chronic lymphocytic leukaemia.

Heinz bodies began to appear in the blood after 3 months feeding in mice fed metanil yellow at 3.0 g/kg body weight and were observed in 80-90% erythrocytes at the end of the experiment.

Blood showed marked anisocytosis, poikilocytosis, stomatocytes, schistocytes and a large number of target cells, all normochromic in character. An
overwhelming preponderance of mature lymphocytes with scant cytoplasm, larger and denser clumps of nuclear chromatin and accentuation of parachromatin were also observed, which were suggestive of chronic lymphocytic leukaemia.

Clinical chemistry: No data

Urinanalysis: No data

Neurobehaviour: No data

Organ weights: No data

Gross pathology: Mice fed metanil yellow at 3000 mg/kg bw showed hepatomegaly and splenomegaly at autopsy.

Histopathology: No data

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw (total dose)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
gross pathology
haematology

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table I Haematological Findings in Male Mice Fed with Metanil Yellow at 0.0 and 3.0 G/Kg Body Weight Concentrations

No.

Experiment

SI Unit

0 mg/Kg bw

3000 mg/Kg bw

1

Total erythrocyte count (TEC)

1012/L

6.13±0.45

3.86 ± 0.70

2

Differential leukocyte count (DLC)

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

109/L

 

 

5.67 ± 0.23

3.28 ± 0.16

0.44 ± 0.02

0.05 ± 0.01

0.00 ± 0.00

 

 

0.44 ± 0.16

9.10±0.44

0.65±0.03

0.01 ± 0.00

0.01 ± 0.00

3

Haemoglobin (Hb)

g/dL

16.29 ± 1.30

13.64 i 0.55

4

Erythrocyte sedimentation

rate (ESR)

mm/h

1.81±0.25

4.18 ± 0.75

5

Mean corpuscular

volume (MCV)

FI

11.77 ± 1.77

139.88±10.00

6

Mean corpuscular

haemoglobin (MCH)

Pg

24.94±2.88

35.41±0.79

7

Heinz bodies

% of erythrocytes

0.00±0.00

89.25±2.50

Mean± Standard error 

 

Table II Haematological Findings in Female Mice Fed with Metanil Yellow at 0.0 and 3.0 G/Kg Body Weight Concentrations

No.

Experiment

SI Unit

0 mg/Kg bw

3000 mg/Kg bw

1

Total erythrocyte count (TEC)

1012/L

5.26±0.37

3.49 ± c0.21

2

Differential leukocyte count (DLC)

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

109/L

 

 

5.14 ± 0.28

3.09 ± 0.19

0.40 ± 0.04

0.04 ± 0.01

0.00 ± 0.00

 

 

0.32 ± 0.11

9.20 ± 0.54

0.69 ± 0.06

0.01 ± 0.00

0.01 ± 0.00

3

Haemoglobin (Hb)

g/dL

13.54 ± 1.10

10.50 ± 1.00

4

Erythrocyte sedimentation

rate (ESR)

mm/h

2.21 ± 0.30

4.31 ± 0.25

5

Mean corpuscular

volume (MCV)

FI

74.10 ± 5.40

151.86 ± 4.20

6

Mean corpuscular

haemoglobin (MCH)

Pg

25.76 ± 2.97

33.23 ± 0.47

7

Heinz bodies

% of erythrocytes

0.00 ± 0.00

81.75 ± 2.25

Mean± Standard error

 

Applicant's summary and conclusion

Conclusions:
The No Observed Adverse Effect level (NOAEL) for the test compound sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate (metanil yellow) is determined to be 500 mg/Kg bw upon repeated exposure for 180 days to male and female albino mice by oral intubation route.
Executive summary:

Chronic oral toxicity study was performed to determine the oral toxic nature of sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate (metanil yellow) upon repeated exposure for 180 days. The test compound was force fed to male and female albino mice by oral intubation at a dose levels of 0, 100, 500 or 3000 mg/Kg bw.

 

The animals were observed for general condition, changes in hematological parameters and gross pathology.

 

General condition was unaffected at dose levels of 100 and 500 mg/Kg bw except at 3000 mgKg bw. No change was observed by the 100 and 500 mg/Kg bw but feeding of this colour at the rate of 3000 mg/kg body weight led to certain changes in the haematological values. Total erythrocyte count (TEC) and Hb had decreased whereas ESR, MCV, and mean corpuscular haemoglobin (MCH) had increased. These facts suggested the occurrence of normochromic macrocytic anaemia. Differential leukocyte count (DLC) showed marked increase in the number of lymphocytes and monocytes and decrease in the number of neutrophils and eosinophils. Heinz bodies were observed in 80-90% erythrocytes. Other haematological values like total leukocyte count (TLC), platelet count (PC), PCV, MCHC, bleeding time (BT) and coagulation time (CT) were normal at 3000 mg/Kg bw.

 

Hence, the No Observed Adverse Effect level (NOAEL) for the test compound metanil yellow is 500 mg/Kg bw.