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 internationally accepted testing guideline.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
GLP compliance:
no
Test type:
standard acute method
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Velaz Prague.
- Weight at study initiation: about 150 g.
- Housing: semparated, in plastic polypropylene cages T4 (supplied by s.p. Velaz Prague).
- Diet: animals were fed with commercial granular food mixture Altromin 1320, supplied by s.p. Velaz Prague. Daily dose of 15 g/animal/day.
- Fasting period before study: the day before of the test, animals were not fed.
- Water: CSN 757111 ad libitum.
- Bedding: wood shavings, from light wood.
- Acclimation period: 1 week.
Cleaning and disinfection of premises menagerie were made at dates determined, according to the standard operating procedures and compliance regime measures.

ENVIRONMENTAL CONDITIONS
- Temperature: controlled temperature at 22 ± 3 °C.
- Humidity: 50 ± 15 %.
- Photoperiod: 12 hrs dark / 12 hrs light, by fluorescent lamp.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20 %
Doses:
3981, 5020, 6310, 7943, 10000 mg/kg
No. of animals per sex per dose:
10 rats x dose
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: immediately after application, ca 30 minutes, after 3 hours post-application. The day after the application the observations were made in the morning and in the afternoon, while in the following days at least once daily. Body weight was measured at the beginning and end of the experiment.
- Necropsy of survivors performed: yes; internal organs were assessed for colour, size, consistency and structure.
- Other examinations performed: appearance of the skin, hair, visible mucous membranes status, mental activity, somatomotor activity, reactivity to stimulus, lacrimation, respiration, digestion, urogenital and circulatory apparatus. Organs and muscles were examined macroscopically. If post-mortem the bladder of animals was full, the urines were analyzed focusing on the detection of proteins, blood sugar, ketones, bilirubin, urobilinogen and pH.
Statistics:
The LD50 is calculated according to the probit method by Blisse.
Data on mortality, the frequency and using the logarithmic dose levels used, were inserted into a computer and analyzed by the PROBIT program, from October 1991 VUOS.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
ca. 6 620.9 mg/kg bw
Based on:
test mat.
Mortality:
3981 mg/kg: 0.0 %
5020 mg/kg: 30.0 %
6310 mg/kg: 30.0 %
7943 mg/kg: 70.0 %
10000 mg/kg: 100.0 %
Clinical signs:
After 3 hours post-application at dose level of 10.00 g/kg the following symptoms were recorded:
- the appearance of skin and hair: stained with excretions.
- nutritional status: cachectic.
- appearance of visible mucous membranes: in the range of physiological norms.
- mental activity: stick to the physiological norm.
- somatomotor activity: rats were upright in a cage. Motor activity was in the physiological norm.
- reactivity and sensibility: in the physiological norm.
- lacrimation: in the normal range.
- circulatory system function: in the physiological norm.
- respiratory system function: in the physiological norm.
- digestive system function: diarrhea.
- urogenital system function: in the physiological norm.

Symptoms recorded at the lower dose level administered, are analogous to those of the maximum dose.
At the dose level of 3.981 g/kg were not recorded symptoms of intoxication, nor macroscopically and patomorphological changes; therefore, it can be considered as no effect dose level.
Other findings:
Autopsies of dead rats administered at dose of 1000 mg/kg revealed:
- External examination of the cadaver
Appearance of hair, skin: hair contaminated with excretions.
Nutritional status: cachectic.
Natural body openings: no macroscopical, patomorphological changes.
Integrity of the body surface: intect.
Postmortality: rigor mortis.

- Organs of head and neck: no macroscopical, patomorphological changes.

- Internal inspection cadaver
Tongue, esophagus, trachea: no macroscopical, patomorphological changes.
Lungs: pink, spongy consistency, airy, without macroscopical, patomorphological changes.
Heart: brownish-red colour, stiffer consistency, without macroscopical, patomorphological changes.

- Abdominal organs
Stomach: acute dilatation, filled with mash content, wall colour brown.
Guts: filled with sparse content, brown-coloured wall
Liver: brownish-red color, stiffer consistency, without macroscopical, patomorphological changes.
Spleen: stiffer consistency, red colour, without macroscopical, patomorphological changes.
Kidney: brownish-red colour, stiffer consistency, without macroscopical, patomorphological changes.
Peritoneum: no macroscopical, patomorphological changes.
Bladder: empty, without macroscopical, patomorphological changes.

- Urine - biochemical tests not established

Symptoms recorded at the lower dose level administered, are analogous to those of the maximum dose.

Any other information on results incl. tables

Overview of the time interval after the onset of various levels of logarithmic dose
Dose
mg/kg
Percent mortality 
Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14
1/2 hr 3 hrs
3981 0 0 0 0 0 0 0 0 0 0 0 0 0 0
5020 0 30 0 0 0 0 0 0 0 0 0 0 0 0
6310 0 20 10 0 0 0 0 0 0 0 0 0 0 0
7943 0 70 0 0 0 0 0 0 0 0 0 0 0 0
10000 0 70 0 0 0 0 0 0 0 0 0 0 0 0

Overview of mortality by sex of experimental rats at different levels
Mortality for each experimental group of tests for acute oral toxicity.
Dose
mg/kg
Number of dead animals
Percentage
Males Females
3981 0 0 0
5020 1 2 30
6310 0 3 30
7943 4 3 70
10000 5 5 100

Body weight

Dose
mg/kg
Males
g/kg
Females
g/kg
Start test End test Difference Start test End test Difference
3981 166.4 173 + 6.6 160 168 + 8
5020 167.6 178.8 + 11.22 154.4 158.8 + 44.4
6310 160.8 171.6 + 10.8 159.6 154.8 - 4.8
7943 156 148.8 - 7.2 163.2 166 - 3.2
1000 162.8 163.2 - 9.6 148.8 148.4 - 0.4

Results

Dose
g/kg
Group Percentage of mortality Log. Dose Probits Exp.
3981 10 0.0 0.59999
5020 10 30.0 0.70070 4.476
6310 10 30.0 0.80003 4.476
7943 10 70.0 0.89998 5.524
10000 10 100.0 1.00000

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU
Conclusions:
Oral LD50: 6620.9 mg/kg (6098.6 - 7187.9 mg/kg).
Executive summary:

The acute toxicological characterization of the substance was determined by Acute toxicity test, according to the OECD guideline 401.

Groups of ten rats (5 males and 5 females) were administrated by oral gavage with 3981, 5020, 6310, 7943, 10000 mg/kg of test material. The LD50 was calculated according to the probit method by Blisse.

Conclusion

The LD50 was calculated to be 6.6209 g/kg (6.0986 - 7.1879 g/kg).