Reaction mass of tetrasodium 7,7'-(carbonyldiimino)bis[4-hydroxy-3-[(2-methyl-4-sulphonatophenyl)azo]naphthalene-2-sulphonate] and tetrasodium 4-[[1-hydroxy-6-[[[[5-hydroxy-6-[(2-methyl-4-sulphonatophenyl)azo]-7-sulphonato-2-naphthyl]amino]carbonyl]amino]-3-sulphonato-2-naphthyl]azo]benzoate and tetrasodium 4,4'-[carbonylbis[imino(1-hydroxy-3-sulphonatonaphthalene-6,2-diyl)azo]]dibenzoate

Endpoint summary

Description of key information

The NOAEL was determined to be 1000 mg/kg bw/day.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

From January 15 to February 28, 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: No GLP, aged study. Meets generally accepted scientific standards, well documented and acceptable for assessment.

Qualifier:

no guideline followed

Deviations:

not applicable

Principles of method if other than guideline:

1000 mg/kg/day of FAT 11'110/N where administrated daily (5 times a week) by gavage over a period of 30 days to male and female rats.

GLP compliance:

no

Limit test:

yes

Species:

rat

Strain:

other: Tif: RAIf (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: approx. 4 weeks
- Weight at study initiation: males: 128 - 129 g, females: 113 - 115 g
- Housing: individual housing in Macrolon cages type 3 with standardized granulated soft wood bedding (Societé Parisienne des sciures Pantin)
- Diet (e.g. ad libitum): pelleted standard diet (Nafag No. 890) ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 1
- Humidity (%): 55 ± 5
- Air changes (per hr): 15 - 17
- Photoperiod (hrs dark / hrs light): 14 / 10

Route of administration:

oral: gavage

Vehicle:

CMC (carboxymethyl cellulose)

Remarks:

2%

Details on oral exposure:

DOSING SOLUTIONS:
daily prepared suspension of the test compound in CMC 2%; control animals received a similar trweatment with CMC 2% without the compound

VEHICLE
- Concentration in vehicle: 10% compound in CMC 2%
- Amount of vehicle (if gavage): 10 ml/kg

Duration of treatment / exposure:

4 weeks and 2 days of the 5th week (30 days)

Frequency of treatment:

5 times a week (in total 22 doses)

Remarks:

Doses / Concentrations:
1000 mg/kg bw/d
Basis:
actual ingested

No. of animals per sex per dose:

20 (10 male and 10 female animals had to undergo a two week recovery period)

Control animals:

yes, concurrent vehicle

Details on study design:

- Post-exposure recovery period in satellite groups: 2 weeks

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: mortality: daily (a.m. and p.m.), symptoms: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Time schedule: weekly

FOOD EFFICIENCY:
- Mean food conversion (g food/kg bw/day) was calculated according to the following formula: MFC = (weekly food consumption (g)) / (midweek body weight (g)) * 1000/7

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: by observation prior to dosing and after the treatment period

HEARING TEST: Yes
- Time schedule for examinations: prior to dosing and after the treatment period (by hand clapping)

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at test day 28
- Anaesthetic used for blood collection: No
- Animals fasted: Yes
- How many animals: 20 randomized rats (5 males, 5 females per group)
- Parameters examined: Hb, RBC, WBC (total count and differential count)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at test day 28
- Animals fasted: Yes
- How many animals: 20 randomized rats (5 males, 5 females per group)
- Parameters cexamined: GOT, GPT, AP

URINALYSIS: Yes
- Time schedule for collection of urine: at test day 28
- Animals fasted: Yes
- Parameters examined: pH, protein, glucose, urobilinogen, urine sediment

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Macroscopical Examination: At the end of the 30 day toxicity study or after an additional recovery period of two weeks the treated and control rats were bled under ether anaesthesia and complete autopsy was carried out. The total weight of each animal was determined and following organs were weighed: liver, kidneys and adrenals. The mean organ weights and mean organ to body weight ratios of each of these organs were calculated for all treated and control groups.

HISTOPATHOLOGY: Yes
Tissue portions of trachea, lungs, liver, pancreas, salivary gland, stomach, small and large intestine, heart, spleen, lymph nodes, thymus, kidneys, urinary bladder, testicles or ovaries, prostate or uterus, pituitary, thyroid, adrenals, skeletal muscle, skin, malMlary glands, bone marrow, eyes, brain, spinal cord , sciatic nerve, oesophagus and all other organs and tissues which showed macroscopical changes were fixed in buffered 10 % formalin.
The fixed tissues samples of liver, kidneys and adrenals were embedded in paramat and cut at 3 - 5 µ. The routine stain was haematoxylin and eosin. Sections from liver, kidneys and adrenals were also stained by Perl's method for·iron.
Additional frozen sections from liver were specifically stained for fat with Sudan Ill. Only the liver, kidneys and adrenals were histologically examined.

Statistics:

For each time point and parameter a uni-variate statistical analysis was conducted. Due to the routine manner of the analysis system parameter free methods were applied. The treated group was compared to the control group in respect of dispersion and displacement (Y. Lepage, Biometrika (1971) 58: pp. 213-217).

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Ophthalmological findings:

no effects observed

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No animal died during the treatment and/or the recovery period. No clinical symptoms and no signs of local and/or systemic toxicity were observed.

BODY WEIGHT AND WEIGHT GAIN
The body weight gain of all treated and control rats was comparable during the test and the recovery period.

FOOD CONSUMPTION
The mean food consumption of all treated animals was comparable to the controls during the 30 days of administration and during the recovery period.

FOOD EFFICIENCY
The mean food conversion of the males and females of the treated group was comparable to the controls.

OPHTHALMOSCOPIC EXAMINATION
Eye examinition did not reveal any ocular changes.

AUDITORY PERCEPTION
No loss of hearing ability was registred.

HAEMATOLOGY
The observed haematological findings between treated rats and controls were unremarkable.

CLINICAL CHEMISTRY
In the assessment of blood chemistry values the findings were unremarkable and comparable to those of the controls.

URINALYSIS
The findings in the urine were unremarkable. Individual rats revealed some degree of physiological proteinuria including those of the control group. This is considered normal in laboratory rats.

ORGAN WEIGHTS
Organ weights and organ to bodyweight ratios revealed no statistically significant differences between treated and control animals.

GROSS PATHOLOGY
No compound related gross anatomical changes were noted.

HISTOPATHOLOGY
All minor changes seen in some control and treated rats were only incidental in nature and not related to the treatment withthe test item.

Dose descriptor:

NOEL

Effect level:

> 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Critical effects observed:

not specified

Conclusions:

It can be concluded from the observations made during the study that 1000 mg/kg/day of the test item produced no observable effects in male and female rats when administrated daily by gavage over a period of 30 days.

Executive summary:

The toxicity of FAT 11'110/N was investigated during a period over 30 day on females and males rats by oral administration.

No observable effects were observed at a dosage of 1000 mg/kg/day.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Two studies are available performed with the same structural analogue. Both were determined to be klimisch 2.

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Studies were performed with analogue substance 1 (please refer to IUCLID section 13).

 

Repeated dose toxicity: oral

The toxicity of the structural analogue was investigated in an oral repeated dose study. Ten Tif: RAIf (SPF) rats per sex were exposed to the test substance dissolved in a 2% carboymethyl cellulose solution via oral gavage. Animals received 1000 mg/kg bw/day, five times per week for a period of five weeks. Animals receiving the vehicle served as controls. Clinical signs, body weight, food intake, food efficiency, ophthalmoscopic examination, auditory perception, hematology, blood chemistry, urinalysis, gross pathology, organ weight and histopathology were investigated. No toxic effects were observed. Based on this information the NOEL was calculated to be 1000 mg/kg bw/day.

Leist et al (1982) published the results of subacute toxicity studies on eight selected colorants, including the structural analogue. These eight colorants were administered as 22 daily doses over a period of 30 days (no weekend dosing) at 1000 mg/kg body weight/day by gavage to 20 female and 20 male rats. Animals treated with the vehicle served as controls. In most cases the treatment period was followed by a 15-day recovery period. Clinical signs, body weight, food intake, hematology, blood chemistry, urinalysis, gross pathology, organ weight and histopathology were investigated. At this dose level all products were tolerated without irreversible signs of toxicity. The survey of data available indicated that the substances investigated exhibited a very low cumulative toxicity. The NOAEL for the structural analogue was therefore determined to be 1000 mg/kg bw/day.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
There are two studies available. Both studies are performed with the same structural analogue. The study with the most detailed information has been selected as key.

Justification for classification or non-classification

No signs of toxicity were observed after repeated exposure to 1000 mg/kg bw in two studies with analogue substance 1. Based on this information classification for oral repeated dose toxicity is not warranted in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.