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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
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
Qualifier:
according to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Version / remarks:
OECD 407 (1981)
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Reactive Red 239

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Remarks:
deionised
Details on oral exposure:
Method of administration:
gavage
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
Test duration: 28 days
Frequency of treatment:
Dosing regime: 7 days/week
No. of animals per sex per dose:
Male: 5 animals at 0 mg/kg bw/day
Male: 5 animals at 40 mg/kg bw/day
Male: 5 animals at 200 mg/kg bw/day
Male: 5 animals at 1000 mg/kg bw/day
Female: 5 animals at 0 mg/kg bw/day
Female: 5 animals at 40 mg/kg bw/day
Female: 5 animals at 200 mg/kg bw/day
Female: 5 animals at 1000 mg/kg bw/day

Examinations

Observations and examinations performed and frequency:
Behaviour and general health were observed in all animals were observed twice a day during the whole experiment and on week-end once a day. Neurolocial effects, eyes, oral mucosa and effects on the teeth growh were observed weekly. Bodyweight were measured at the start of the experiment and then twice a week.
Statistics:
statistical significant differences were analysed by:
Body Weight, hematological parameter, clinical-chemical parameter, Albumin and globulin values, absolute and relative organs weights
PH wert and weight of Urine.

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:
effects observed, treatment-related
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
reduction in the creatine value and alpha-1-globulin values occurred
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
coloured
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
elevaton of he liver and kidney weights in high dose group
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
Clinical observations:
No mortalities occurred. In the high-dose group, red discoloration of the urine was observed as from the 4th day of testing and red discoloration of the hairless parts of the body as from the 20th day of testing.

Laboratory findings: Haematologically, there were no differences from the controls detected. Clinico-chemically, changes in the form of a significant reduction in the creatine values and the alpha 1-globulin values occurred in particular among the females from the high-dose group.

Effects in organs: In addition, the adrenal weights (absolute and relative) were clearly elevated among the males from the high-dose group as were the relative liver weights of the females from the medium and high-dose groups and the relative kidney weights in the high-dose group. Macroscopically, a slight red discoloration of kidneys was diagnosed among the females as from 40 mg/kg and among the males as from 200 mg/kg. In the high-dose group, all of the animals exhibited clear red discoloration of the kidney, the stomach and the connective tissue and, among the males, of the testes and of the intestine too. Pathohistologically, dose-dependent deposits of the test substance were found in the tubular epithelium of the kidneys of the animals from the medium and high-dose groups, something which was interpreted as reabsorption of dye particles and lysosomal accumulation. All of the other findings were not dose-related and correspond to the spontaneous pathological picture for the age group of the species.

Effect levels

open allclose all
Dose descriptor:
NOEL
Effect level:
200 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effects observed
Dose descriptor:
NOAEL
Effect level:
>= 1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: dicolouration of inner organs and tissues

Target system / organ toxicity

Critical effects observed:
no

Applicant's summary and conclusion

Conclusions:
The 'no observed effect level' (NOEL) in this study is 200 mg/kg bw/day. Howerer, clear signs of toxic effects were also not seen in the 1000 mg/kg bw group, hence, the 'no observed adverse effect level' (NOAEL) is considered to be 1000 mg/kg bw/day.
Executive summary:

Reactive Red 239 was administered overa period of 29 days, SPF-Wistar rats at dose levels of 0, 40, 200 and 1000 mg/kg body weight per day orally by gavage (total 28 applications, 7 days a week).In all experimental groups, the behavior and general health were checked twice daily, on weekends and holidays once daily. Body weight gain and food consumption were determined twice a week; water consumption weekly. Haematological, clinical chemistry and urinalysis tests were performed at the end of the study.

At necropsy, the animals were examined macroscopically for organ changes; the major organs were weighed and relative organ weights were calculated. The most important organs of these animals were collected and histological specimens were prepared and examined for microscopic changes. Body weights, hematology and clinical chemistry parameters, albumin and globulin values, urinalysis (pH, density) and the absolute and relative organ weights were statistically tested for differences compared to the control groups.

The 28-day administration of 1000 mg/kg bw Reactive Red 239 resulted in no compound-related impairment of general health status or body weight gain. Feed and water consumption were also unaffected by the test substance.

The hematological investigations showed no evidence of compound-related toxicity.
The evaluation of the clinical-chemical parameters resulted in lower creatinine values in females of the 1000 mg/kg bw-group. All other changes were within the normal range of the used rat strain and are therefore not related to the administration of the test substance administration. In the animals of the 200 and 1000 mg/kg bw-groups, the urine was brown-yellow to red discolored. Otherwise, the urinalysis was normal.

In male animals at 1000 mg/kg bw, the adrenal weights were increased. In females at the 200 and 1000 mg/kg bw, increases in relative liver weights and at 1000 mg/kg bw increases in relative kidney weights were observed.

The macroscopic examinations revealed reddish discoloration of various tissues and organs. The affected organs were the kidneys of the 40 mg/kg bw-group females, 200 mg/kg bw-group males, and 1000 mg/kg bw-group rats, as well as the connective and stomach tissue at 1000 mg/kg bw. In addition, in males the testes and colon were stained red.

Microscopically, dose-dependent increase in deposits of the substance in the proximal tubulesof the kidneys were found in the top two dose groups (200 and 1000 mg/kg bw), which is interpreted to be a sign of reabsorption of the dye resulting in intra-cellular lysosomal storage. No other substance-related changes were observed.

In summary, in this 29-day subacute toxicity test (28 applications in 29 days)the administreation of Reactive Red 239 resulted in a dose-dependent incorporation of the substance in the proximal tubule of the kidneys without causing changes in renal morphology in animals of the 200 and 1000 mg/kg bw groups.Since the clinical chemistry parameters revealed also no evidence of impaired renal function in the 1000 mg/kg bw-group,a toxic effect is unlikely in this dose group.

Consequently, the 'no observed effect level' (NOEL) in this study is 200 mg/kg bw/day. Howerer, clear signs of toxic effects were also not seen in the 1000 mg/kg bw group, which was defined as the 'no observed adverse effect level' (NOAEL).