Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 12 JULY 2018 to 11 DECEMBER 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018
Report Date:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Version / remarks:
OECD Nº423 DEcember 2001.
OECD Guideline for testing of chemicals Acute Oral Toxicity - Acute Toxic Class Method.
Deviations:
yes
Remarks:
Relative humidity/temperature in the room where the animals were housed was sporadically below/above the optimal range during the study period. This deviation was not considered to have any impact on the interpretation of the study results.
GLP compliance:
yes (incl. certificate)
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder

Test animals

Species:
rat
Strain:
Sprague-Dawley
Remarks:
Colour: albino
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Rationale for alternative/additional species to rat (if applicable): Rats are a suitable rodent species, acceptable to regulatory authorities as
indicators of potential hazards, and for which extensive background data are
available. Literature surveys of conventional LD50 tests showed that, although
there is little difference in sensitivity between the sexes, in those cases where
differences were observed females are generally slightly more sensitive
- Source: Envigo RMS Spain S.L.
- Females (if applicable) nulliparous and non-pregnant: [yes/no]: YES
- Rationale for use of males (if applicable)
- Age at study initiation: 8 weeks old
- Weight at study initiation: mean 185.277 g (183.08 (1), 189.56 (2), 183.19 (3))
- Fasting period before study: Animals were fasted overnight before administration and approx. 3-4h afterwards
- Housing:3 animals per cage after distribution. Animals were kept in groups. Enrichment devices (nesting material, tubes, chew blocks) were provided as a default husbandry practice,
- Historical data: Prior to inclusion in the study, animals were subjected to individual veterinary
examination. Only animals without clear visible sign of illness that may interfere
with the study were included.
- Diet (e.g. ad libitum): Ad libitum except for an overnight fast prior to dosing and approx. 3-4h afterwards
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days. Animals were randomly distributed by means of the body weight
stratification method. 20% of the mean body weight (for the first defined dose at
the beginning of the study and further dose steps).
- Microbiological status when known
- Method of randomisation in assigning animals to test and control groups : Animals were randomly distributed by means of the body weight stratification method.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25ºC
- Humidity (%): 30-70%
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12 hours

IN-LIFE DATES: From: (1) 16/07/2018 (2) 18/07/2018 To: (1) 30/07/2018 (2) 01/08/2018

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
methylcellulose
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 30 mg/ml
- Amount of vehicle (if gavage): The required amount of test item was weighed in an appropriate container and the corresponding volume of vehicle was added so that a final concentration
- Justification for choice of vehicle: Suitable for gavaging the rats.
- Lot/batch no. (if required): SLBT8618
- Purity: 0.5% w/v

MAXIMUM DOSE VOLUME APPLIED: 300 mg/Kg

DOSAGE PREPARATION (if unusual):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:
Doses:
30 mg/kg bw
No. of animals per sex per dose:
2 controls, 3 in dose groups.
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30 minutes, 2h, 4h and daily thereafter during the 14-day observation period.
- Necropsy of survivors performed: yes
- Clinical signs including body weight: Any visible clinical signs, discomfort and mortality were recorded. Clinical signs included changes in skin, fur, eyes and mucous membranes. Alterations in respiratory pattern, behaviour, posture, response to handling and the presence of abnormal movements were reported as well. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other: Body weight was recorded once during the acclimatisation period for group distribution, immediately before administration for dosing purposes (from 3-4h fasted animals), weekly thereafter and prior to sacrifice.
Statistics:
No statistical analysis was performed: all the controls survived.

Results and discussion

Preliminary study:
Group A and Group B were treated with a dose level of 300 mg/kg and none of the treated animals shown a test item-related mortality.
In standard conditions a next group with 3 more animals would be treated at the maximum dose level, 2000 mg/kg, according to the OECD Guideline 423, but due to problems with the insolubility of the test item at the maximum dose level it was not possible to check the mortality of a third group at 2000 mg/kg.
Therefore, it can be concluded that, according to the results obtained in this study and under the assayed experimental conditions, the test item could be ranked in the Category 4 (LD50 cut-off value of 300 < LD50 < 2000 mg/kg of body weight), of the Globally Harmonised Classification System for Chemical Substances and Mixtures.
Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
ca. 300 mg/kg bw
Based on:
test mat.
Mortality:
Group A and Group B were treated with a dose level of 300 mg/kg and none of the treated animals shown a
test item-related mortality.
Clinical signs:
Any visible clinical signs, discomfort and mortality were recorded. Clinical signs included changes in skin, fur, eyes and mucous membranes. Alterations in respiratory pattern, behaviour, posture, response to handling and the presence of abnormal movements were reported as well. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma.
Body weight:
All the animals gained weight.
Gross pathology:
There weren't reatment related effects.
Other findings:
- Organ weights: Any remaining protocol tissues, which have been examined, have no visible lesions.
- Histopathology: Any remaining protocol tissues, which have been examined, have no visible lesions.
- Potential target organs: Any remaining protocol tissues, which have been examined, have no visible lesions.
- Other observations: Any remaining protocol tissues, which have been examined, have no visible lesions.

Applicant's summary and conclusion

Interpretation of results:
Category 4 based on GHS criteria
Remarks:
It can be concluded that, according to the results obtained in this study and under the assayed experimental conditions, the test item could be ranked in the Category 4 (LD50 cut-off value of 300 < LD50 < 2000 mg/kg of body weight), of the Globally Harmonised Classification System for Chemical Substances and Mixtures.
Conclusions:
The test item could be ranked in the Category 4.