Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
04-20 February 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Modern guideline study performed in GLP certified laboratory

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

Materials and methods

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

Test material

Constituent 1
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Batch No.: RW 2013-10-22
Form: Powder
Color: Orange
Expiring date: 22.10.2018
Storage conditions: room temperature
Correction factor: 1.05

Test animals

Species:
rat
Strain:
other: Crl:(WI)BR
Sex:
female
Details on test animals or test system and environmental conditions:
Species and strain: Crl:(WI)BR rats
Source: TOXI COOP ZRT. Cserkesz u. 90.
1103 Budapest, Hungary
Hygienic level at arrival: SPF
Hygienic level during the study: Good conventional
Justification of strain: The Wistar rats as a rodent is one of the standard species of acute toxicity studies
Number of animals: 3 animals/group
Sex: Female, nulliparous and non pregnant animals
Age of animals: Young adult rat, 9 weeks old in first and second step
Body weight range
at starting (first step): 180 - 186 g
Body weight range
at starting (second step): 180 - 184 g
Acclimatization time: 12 days in first step and 13 days in second step

Husbandry:

Animal health: Only healthy animals were used for the study. Health status was certified by the study director.
Room: 5/II (E building)
Housing: Group caging (3 animals/cage)
Cage type: Type II polypropylene/polycarbonate.
Bedding: Laboratory bedding.
Light: Artificial light, from 6 a.m. to 6 p.m.
Temperature: 22 ± 3 °C
Relative humidity: 30 - 70 %
Ventilation: 10-15 air exchanges/hour by central air-condition system.

The temperature and relative humidity were recorded daily during the study.
Before housing the animals the microbiological status of the room was checked.

Food and Water Supply:

Animals received ssniff® SM R/M-Z+H complete diet for rats and mice produced by ssniff Spezialdiäten GmbH, D-59494 Soest Germany and tap water from municipal supply, as for human consumption from bottle ad libitum.

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. Copies of the relevant Certificates of Analysis are maintained in Toxi-Coop Zrt.’s archive.

Identification:

The individual identification was performed by numbers on the tail written by a permanent marker. The numbers were given on the basis of Toxi-Coop Zrt.'s master file, for each animal allocated to the study.
The boxes were identified by cards, holding information about study number, sex, dose group, cage number and individual animal numbers.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Sunflower oil (Helianthi annui ol. raffinat.)
Details on oral exposure:
A single oral administration - followed by a fourteen-day observation period - was performed by gavage. The day before treatment the animals were fasted. The food but not water was withheld overnight. Animals were weighed before the application and the food was given back 3 hours after the treatment.
Doses:
Starting dose was selected on the basis of the available information about the test item.
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 Annex 2d of OECD Guideline No. 423 (presented in Appendix VII) was met.
No. of animals per sex per dose:
3 females
Control animals:
no
Details on study design:
step 1 2000 mg/kg bw 3 female rats
step 2 2000 mg/kg bw 3 female rats
Statistics:
no statistical evaluation was necessary

Results and discussion

Mortality:
No death occurred at 2000 mg/kg bw single oral dose of Yellow LF 6881. All female rats in step 1 and step 2 survived until the end of the 14-day observation period.
Clinical signs:
In group 1 (Step 1) treated with 2000 mg/kg bw dose clinical sign of reaction comprised of brownish-red coloured faeces (6 cases of 57 observations). This symptom (score +4; +3) was detected in all animals on Day 1 and on Day 2.

In group 2 (Step 2) treated with 2000 mg/kg bw dose clinical sign of reaction comprised of brownish-red coloured faeces (6 cases of 57 observations). This symptom (score +4; +2) was detected in all animals on Day 1 and on Day 2.
Body weight:
The mean body weight of the all animals corresponded to their species and age throughout the study.
Gross pathology:
All animals survived until the scheduled necropsy on Day 15.
Slight hydrometra was observed in female No.: 8 of the group 1 and in female No.: 11 of the group 2. It 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.

Any other information on results incl. tables

Dose
(mg/kg bw)

Mortality
(dead/treated)

LD50
(mg/kg bw)

GHS
category

2000

0/6

above 2000

5

 

Applicant's summary and conclusion

Interpretation of results:
Category 5 based on GHS criteria
Remarks:
Migrated information
Conclusions:
The method used, was not intended for the precise calculation of a precise LD50 value.
The test item was ranked into Class 5 of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423.
Executive summary:

An 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, therefore 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 Annex 2d 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.

Lethality, Clinical symptoms and Body weight:

No death occurred during the study. All rats dosed at 2000 mg/kg bw test item Yellow LF 6881 survived until the end of the 14-day observation period. In the first step, brownish-red coloured faeces was observed in animals between Day 1 and Day 2. In the second step, brownish-red coloured faeces was observed in animals between Day 1 and Day 2. The observed clinical sign as brownish-red coloured faeces was not related to the systemic toxic effect of the test item, but this alteration was connected with the physical property of the test item. The body weight development was undisturbed in all animals.

Gross pathology:

Six animals were sacrificed as scheduled during the study. All organs of all experimental animals proved to be free of treatment related gross pathological changes.

Evaluation:

The method used is not intended to allow the calculation of a precise LD50 value. The test item was ranked into Class 5 of Globally Harmonized Classification System (GHS) described in the OECD Guideline No. 423.