Reaction mass of lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-[(4-{[2-(sulfonatooxy)ethyl]sulfonyl}phenyl)diazenyl]naphthalene-2,7-disulfonate and lithium sodium hydrogen 4-amino-6-({5-[(5-chloro-2,6-difluoropyrimidin-4-yl)amino]-2-sulfonatophenyl}diazenyl)-5-hydroxy-3-{[4-(vinylsulfonyl)phenyl]diazenyl}naphthalene-2,7-disulfonate

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18-07-1986 to 15-08-1986

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Lithium sodium hydrogen 4-amino-6-(5-(5-chloro-2,6-difluoropyrimidin-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate

EC Number:

401-560-2

EC Name:

Lithium sodium hydrogen 4-amino-6-(5-(5-chloro-2,6-difluoropyrimidin-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate

Cas Number:

108624-00-6

Molecular formula:

C28H(21-x-y)ClF2Li(x)N8Na(y)O16S5

IUPAC Name:

Lithium sodium hydrogen-4-amino-6-(5-(5-chloro-2,6-difluoropyrimidine-4-ylamino)-2-sulfonatophenylazo)-5-hydroxy-3-(4-(2-(sulfonatooxy)ethylsulfonyl)phenylazo)naphthalene-2,7-disulfonate

Test material form:

solid: particulate/powder

Details on test material:

Reactive Blue FC 05717

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: males - 126 g (115-137 g), females - 115 g (108-128 g)
- Housing: in Makrolon-Kafigen Type II cages
- Diet (e.g. ad libitum): Altromin 1324 pellets
- Water (e.g. ad libitum): ad libitum

DETAILS OF FOOD AND WATER QUALITY:

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-22°C
- Humidity (%): 50%
- Photoperiod (hrs dark / hrs light): 12:12

IN-LIFE DATES: 18-07-1986 to 15-08-1986

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Due to the good dispersibility of reactive blue FC 05717 in water in the concentration range used, a homogeneous distribution of the test suspension was obtained in the formulations. The test substance was administered to the rats via oral gavage. The test substance suspension was found to be stable for 48 h once prepared.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

All the test concentrations were analytically verified at least once during the study.

Duration of treatment / exposure:

28 d

Frequency of treatment:

Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

40 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

200 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

six rats per sex per dose

Control animals:

yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:

The animals were regularly observed, weighed and the feed intake determined.

Sacrifice and pathology:

At the end of treatment, hematological and clinical-chemical investigations were carried out as well as organ weights (testes, liver, kidneys, adrenal glands). Heart, testes, liver, lung, spleen, kidney, adrenal, and other macroscopically abnormally altered organs/tissues were histopathologically examined.

Results and discussion

Results of examinations

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

Water consumption and compound intake (if drinking water study):

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

The relative liver weight in 1000 mg/kg/day females were significantly increased compared to controls. Although the absolute liver weight in these females were also elevated but there was no statistical significance.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

All animals at 1000 mg/kg/day presented a blue coloration of the visible connective tissue, particularly in the skin area, dark blue kidneys and bluish gastrointestinal contents. Also, in the females of the highest dose, approximately 10% higher liver weight was observed which was interpreted by the authors as a sign of a non-specific adaptation due to increased metabolic activity.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Effect levels

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no treatment-related systemic effects observed

Target system / organ toxicity

Key result

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the systemic NOAEL of the test substance in rats was considered to be 1000 mg/kg/day.

Executive summary:

A study was conducted to determine the oral repeated dose toxicity of the test substance in rats according to OECD Guideline 407 and EU method B.7, in compliance with GLP. Male and female Wistar rats (6/sex/dose) were administered the following concentrations via oral gavage (suspended in water, at a constant volume of 10 mL/kg): 0, 40, 200 and 1000 mg/kg bw/day. Mortality, clinical signs, body weight and food consumption were recorded daily. At necropsy, blood samples were collected for haematology and clinical chemistry investigations. Also, a gross macroscopical examination was conducted, and organs were weighed. Routine histopathological examination was performed on the major organs such as heart, liver, kidneys, testes, adrenals, spleen, lungs etc. No treatment-related clinical signs of toxicity were observed. There were no treatment-related adverse effects on the body weight, food consumption, hematological and clinical biochemistry examinations. All animals at 1000 mg/kg bw/day presented a blue coloration of the visible connective tissue, particularly in the skin area, dark blue kidneys and bluish gastrointestinal contents. This was attributed to the inherent colour of the test substance. Also, in the females of the highest dose, approximately 10% higher liver weight was observed which was interpreted by the authors as a sign of a non-specific adaptation due to increased metabolic activity. Under the study conditions, the systemic NOAEL of the test substance in rats was considered to be 1000 mg/kg bw/day (Schmidt and Sander, 1988).