Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
31-JAN-2007 to 12-JUN-2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2007
Report Date:
2007

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)
Deviations:
no
GLP compliance:
yes
Test type:
acute toxic class method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
ACUTE ORAL TOXICITY STUDY
- Name of test material (as cited in study report): Addotto MOVE 3
- Substance type: monoconstituent 
- Physical state: colourless clear free-flowing liquid
- Analytical purity, impurities, purity test date: no data available
- Lot/batch No.: DIST.05/06
- Expiration date of the lot/batch: 31-DEC-2010
- Storage condition of test material: ambient condition

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Italy S.r.l. / 33049 San Pietro al Natisone (UD) / ITALY
- Age at study initiation: 6 to 8 weeks
- Weight at study initiation: 176 to 200 g; the body weight of each individual was within 20% of the mean of the group.
- Fasting period before study: overnight fast prior to dosing and a period of approximately 4 hrs after dosing
- Housing: in groups of 3 animals, in polycarbonate cages (59 x 38.5 x 20 cm) with stainless steel mesh lid and floor
- Diet: ad libitum, commercial laboratory rodent diet (4 RF 18, Mucedola S.r.l. / 20019 Settimo Milanese (MI) / ITALY)
- Water: ad libitum, drinking water
- Acclimation period: at least 5 days before the start of the test

ENVIRONMENTAL CONDITIONS
- Temperature: 22 ± 2°C
- Humidity: 55 ± 15%
- Air changes: 15 to 20 air changes per hour
- Photoperiod: 12 hrs dark / 12 hrs light

IN-LIFE DATES: From 26-JAN-2007 to 06-MAR-2007

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
as 0.5% aqueous solution
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200 mg/mL
- Amount of vehicle: 10 mL/kg
- Justification for choice of vehicle, lot/batch no., purity: no data available

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

CLASS METHOD
- Rationale for the selection of the starting dose: a single group of 3 female animals was dosed at a level of 2000 mg/kg (step 1). No mortality occurred and three additional females were subsequently dosed at the same dose level (step 2).
Doses:
2000 mg/kg
No. of animals per sex per dose:
3 per group (2 groups treated, for a total of 6 animals per dose)
Control animals:
no
Details on study design:
A single group of 3 female animals was dosed first. At the end of the observation period, an additional group was treated at the same dose level.

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: throughout the study all animals were checked twice daily for mortality and morbidity. Animals were observed for clinical signs immediately upon dosing, approximately 30 min, 2 and 4 hrs after dosing and daily thereafter for a total of 14 days. All animals were weighed at allocation to the study (day -1), immediately prior to dosing (day 1) and on days 2, 8 and 15.
- Necropsy of survivors performed: yes; all animals were killed on day 15 by carbon dioxide narcosis. Animals were subjected to a gross necropsy examination for both external and internal abnormalities. The cranial, thoracic and abnormal cavities were opened to allow examination of their contents. Larger organs were sectioned. Both the stomach and representative sections of the gastro-intestinal tract were opened for examination of the mucosal surfaces.

Results and discussion

Effect levels
Key result
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No death occurred following dosing at 2000 mg/kg in each group of female animals (steps 1 and 2).
Clinical signs:
The only significant clinical sign observed in the first group of animals treated with 2000 mg/kg (step 1) was the presence of soft faeces in the cage tray on Day 2. Recovery from this sign occurred by Day 3. Hair loss of the dorsum was detected in 1/3 female from days 2 to 10, in 2/3 females on day 11 and in 3/3 females from days 12 to 15. This sign had no toxicological significance.
Piloerection was observed in the 3 animals from the second group approximately 2 hours after dosing. Presence of soft faeces in the cage tray was recorded on Day 2. Recovery was completed by Day 3.
Body weight:
Changes in body weight were within the expected range for this strain and age of animals.
Gross pathology:
No abnormalities were observed in any animal at the necropsy examination performed at termination of the study.

Any other information on results incl. tables

Table 1: Body weight and body weight changes (g) - mean data

   

Day -1

(pre-test phase)

Day 1

(dosing phase)

Day 2 Day 8 Day 15 
 Step 1 Mean body weight   219.0 203.7 218.0 233.0 240.3
  Standard deviation (SD) 5.3 3.2 2.0  5.0  7.2 
  Mean body weight change - - +14.3  +29.3  +36.7 
  SD 3.1  2.3  4.2 
 Step 2 Mean body weight  230.3 213.7  230.7  239.3  246.0 
  SD 9.1 11.2  11.0  12.9  13.9 
  Mean body weight change +17.0  +25.7  +32.3 
  SD 3.0  2.1  3.1 

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In conclusion, these results indicated that the test item had no significant toxic effect on the rat following oral administration of a single dose at a level of 2000 mg/kg. The lack of mortality demonstrated the LD50 to be greater than 2000 mg/kg body weight.
Executive summary:

The acute toxicity of the test item was investigated after oral administration (10 mL/kg in 0.5% aqueous solution of carboxymethyl cellulose) of a single dose to fasted Sprague-Dawley rats, followed by a 14-day observation. The method was in accordance with EU method B.1 tris and OECD guideline 423 and in compliance with good laboratory practices (GLP).

A single group of 3 female animals was dosed at a level of 2000 mg/kg and observed for a period of 14 days (step 1). No mortality occurred. The only significant clinical sign observed in the first group of animals treated at 2000 mg/kg (step 1) was the presence of soft faeces in the cage tray on Day 2. Recovery was completed by Day 3.

Three additional female animals were then dosed at the same dose level (2000 mg/kg) and observed for a period of 14 days (step 2). No mortality occurred. Piloerection was observed in the 3 animals approximately 2 hours after dosing. Presence of soft faeces in the cage tray was recorded on Day 2. Recovery was completed by Day 3.

Changes in body weight were within the expected range for this strain and age of animals. Animals were killed at the end of the observation period and were subjected to necropsy examination. No abnormalities were observed at necropsy examination at termination of the study.

These results indicated that the test item had no significant toxic effect on the rat following oral administration of a single dose at a level of 2000 mg/kg. The lack of mortality demonstrated the LD50 to be greater than 2000 mg/kg body weight. Therefore, the test material does not meet the classification criteria of EC Regulation No. 1272/2008 (CLP / EU GHS) and UN GHS for acute toxicity.