Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted according to OECD Guideline 423 and GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

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

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: solid
Details on test material:
Name of test item: NM01
Test item No: 13/0143-2
Batch ID: 492-14-01
Purity: The identity was confirmed. HPLC-analysis revealed: 99.08 area % (190 nm) and 98.17 area-% (200 nm)
Homogeneity: The test item was homogenous by visual inspection
Storage stability: The stability of the test item under storage conditions over the study period was guaranteed by the sponsor, and the sponsor holds this responsibility.
Storage conditions: Room temperature, dry storage
Physical state/color: solid, white

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
Strain: Crl:WI (Han) SPF
Age on day 0: Young adult females (approx. 10 – 11 weeks)
Supplier: Charles River Wiga GmbH, Germany
Arrival in testing facility: Acclimatization period of at least 5 days before the beginning of the experimental phase
Identification: cage cards and tail marking
Body weight on day 0: Animals of comparable weight (± 20% of the mean weight)

Room temperature/relative humidity: The animals were housed in fully air-conditioned rooms. Central air-conditioning guaranteed a range of 22°C ± 3°C for temperature and of 30 – 70% for relative humidity. There were no deviations from these ranges, which influenced the results of the study.
Air changes per hour: approx. 10
Day/night rhythm: 12h/12h
Type of cage: Makrolon cage, type III
Number of animals per cage: single housing
Feeding: VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany
Drinking water: ad libitum
Bedding: H 15005-29; Ssniff, Spezialitäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany)
Enrichment: Wooden gnawing blocks (Type NGM E-022) ; ABEDD® LAB & VET Service GmbH, Hasnerstraße 84/6; 1160 Wien – Austria

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5% in deionized water
Details on oral exposure:
Test item preparation: The test item preparations were produced for each test group shortly before administration by stirring with a high speed homogenizer (Ultra-Turrax) and a magnetic stirrer. Additionally, the homogeneity of the test item preparations during application was ensured by stirring with a magnetic stirrer.
Administration volume: 10 mL/kg bw
Concentration in vehicle: 200 mg/mL (2000 mg/kg bw group), 30 mg/mL (300 mg/kg bw group)
Reason for selection of vehicle: A good homogeneity in water could not be guaranteed, because the test item preparation was a suspension. Therefore, a 0.5% solution of CMC in deionized water was applied.
Form of application: Suspension
Doses:
2000 and 300 mg/kg bw
No. of animals per sex per dose:
3
Control animals:
no
Details on study design:
Fasting period: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
Time of day of administration: In the morning
Observation period: 14 days
Selection of doses: A starting dose of 300 mg/kg bw was chosen in the first step with 3 animals. As no mortality occurred, 2000 mg/kg were administered to another group of 3 animals in the second step. Because no mortality occurred, 2000 mg/kg bw were administered to another group of 3 animals in the third step.
Body weight determination: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation.
Clinical observations: Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter.
Mortality: A check for any dead or moribund animals was made at least once each workday; these records are archived by Bioassay.
Pathology: Necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations.
Histology: No histological examinations were performed.

Results and discussion

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No mortality occurred in all test groups.
Clinical signs:
Clinical signs in the first 2000 mg/kg test group revealed in all animals impaired general state and piloerection at hour 2 after administration only.
No clinical signs were observed during clinical examination neither in the second 2000 mg/kg test group nor in the single 300 mg/kg test group.
Body weight:
The mean body weight of the surviving animals increased within the normal range throughout the study period with one exception in the 300 mg/kg bw test group. One animal showed stagnation of body weight in the second week.
This effect is observed at times in the rat strain used, because in the required age range the female animals have already reached the phase of slow growth. The observed stagnation in the single animal is considered to be unspecific.
Gross pathology:
There were no macroscopic pathological findings in the animals sacrificed at the end of the observation period (9 females).

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of this study the median lethal dose of NM01 after oral administration was found to be greater than 2000 mg/kg bw in rats.
Executive summary:

In an acute oral toxicity study performed according to the Acute Toxic Class method, doses of 2000 and 300 mg/kg bw of the test item NM01 (preparations in 0.5% CMC-solution) were administered by gavage to three test groups of three fasted Wistar rats each (2000 mg/kg bw in 6 females, 300 mg/kg bw in 3 females). Clinical signs were observed in the first test group receiving 2000 mg/kg bw only (impaired general state and piloerection in all animals). No mortality occurred in any of the dose groups. There were no macroscopic pathological findings in all animals sacrificed at the end of the observation period. The mean body weight of the surviving animals increased within the normal range throughout the study period with one exception in the 300 mg/kg bw test group. One animal showed stagnation of body weight in the second week, which was, however, considered an unspecific effect.