Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From July 18th to August 03rd, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
17th December 2001
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Test type:
acute toxic class method
Limit test:
yes

Test material

Constituent 1
Reference substance name:
Acid Red 143
IUPAC Name:
Acid Red 143

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Toxi-coop ZRT, Budapest, Hungary.
- Females: nulliparous and non pregnant animals.
- Age at study initiation: young adult rat, 9-10 weeks old in first and second step.
- Weight at study initiation: first step 203 - 205 g; second step 209 - 221 g.
- Fasting period before study: the day before treatment the animals were fasted. The food but not water was withheld overnight.
- Housing: 3 animals/cage; type III. polypropylene/polycarbonate.
- Diet: animals received ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, ad libitum.
- Water: animals received tap water from municipal supply, as for human consumption from bottle, ad libitum.
- Hygienic level at arrival: SPF
- Acclimation period: 12 days in first step and 13 days in second step.

DETAILS OF FOOD AND WATER QUALITY
The diet and drinking water are periodically analysed and are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 3 °C
- Relative humidity: 30 - 70 %
- Air changes: above 10 air exchanges/hour by central air-condition system.
- Photoperiod: 12 hours daily, from 6.00 a.m. to 6.00 p.m.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
- Concentration: the test item was applied in a concentration of 200 mg/ml
- Volume: concentration of formulations was adjusted to maintain a treatment volume of 10 ml/kg bw.
- Formulation preparation: formulations were prepared just before the administration and were stirred continuously during the treatment.
- Correction factor: 1.28; the correction factor was taken into consideration in the course of the making of solution.
Doses:
2000 mg/kg bw, both first and second step
No. of animals per sex per dose:
3 animals per group per step
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days.
- Mortality: animals were observed individually after dosing at least once during the first 30 minutes, then 1 h, 2 h, 3 h, 4 h after the treatment and twice each day for 14 days thereafter.
- Generla health state: individual observations were performed on the skin and fur, eyes and mucous membranes and also respiratory, circulatory, autonomic and central nervous system, somatomotor activity and behaviour pattern. Particular attention was directed to observation of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Body weighing: the body weights were recorded on day 0 (just before the treatment), on day 7 and on day 15 with a precision of 1 g.
- Necropsy of survivors performed: at the end of the observation period all rats were sacrificed under isofluran anaesthesia. After examination of the external appearance, the cranial, thoracic and abdominal cavities were opened and the appearance of the tissues and organs was observed, and any abnormality was recorded with details of its location, colour, shape and size.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
not determinable due to absence of adverse toxic effects
Mortality:
No death occurred at 2000 mg/kg bw single oral dose of substance. All female rats in step 1 and step 2 survived until the end of the 14-day observation period.
Clinical signs:
In group 1 treated with 2000 mg/kg bw dose clinical sign of reaction comprised of red faeces (9 cases of 57 observations) and diarrhoea (9/57). Red faeces (score +3) and diarrhoea (score +2, +4) were observed in all animals on the treatment day between 1 and 4 hours after the treatment.
In group 2 treated with 2000 mg/kg bw dose clinical sign of reaction comprised of red faeces (12 cases of 57 observations) and diarrhoea (12/57). Red faeces (score +3) and diarrhoea (score +1,+2, +3, +4) were observed in all animals on the treatment day between 1 and 4 hours after the treatment.
Body weight:
The mean body weight of animals treated with 2000 mg/kg bw dose corresponded to their species and age throughout the study.
Gross pathology:
Autopsy revealed no treatment related pathological changes.
All animals treated with 2000 mg/kg bw dose survived until the scheduled necropsy on Day 15. Severe hydrometra was detected in two animals of group 2. Moderate hydrometra was recorded in one animal of group 1. Slight hydrometra was found in one animal of group 1. Hydrometra is physiological finding and connected to the cycle of the animal. No pathological changes were found related to the effect of the test item during the macroscopic examination of animals treated with 2000 mg/kg bw dose.

Applicant's summary and conclusion

Interpretation of results:
other: not classified, according to the CLP Reguation (EC) No 1272/2008
Conclusions:
LD50 > 2000 mg/kg bw (females)
Executive summary:

The acute toxic class method was carried out involving a stepwise procedure with the use of 2000 mg/kg bw as the starting dose in three female rats. No animal died in the first step at 2000 mg/kg bw dose level, so treatment with 2000 mg/kg bw was repeated on further three female rats. No animal died in the second step, too, so the test was finished, the stopping criteria of OECD Guideline No. 423 was met. Animals were weighed, observed for lethality and toxic symptoms for 14 days after the treatment. Gross pathological examination was carried out 15th day after the treatment.

No lethality was noted at single oral dose of 2000 mg/kg bw. In first step, disturbance of the autonomic functions (diarrhoea) was observed in animals on the treatment day between 1 and 4 hours after the treatment. Red faces detected in animals on the treatment day between 1 and 4 hours after the treatment was connected with the physical property of test item.

In second step, disturbance of the autonomic functions (diarrhoea) was observed in animals on the treatment day between 1 and 4 hours after the treatment. Red faces detected in animals on the treatment day between 1 and 4 hours after the treatment was connected with the physical property of test item.

The body weight development was undisturbed in all animals.

All organs of the animals treated with 2000 mg/kg bw proved to be free of treatment related gross pathological changes.

Conclusion

LD50 > 2000 mg/kg bw (females)