Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from August 21, 2007 to Septamber 20, 2007
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study was carried out in accordance with internationally valid GLP principles.

Data source

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

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.3 (Acute Toxicity (Dermal))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Humic acids, sodium salts
- Molecular formula: not known - UVCB substance
- Molecular weight: not known - UVCB substance
- Substance type: technical product
- Physical state: solid
- Lot/batch No.: 9. 5. 2007/R
- Expiration date of the lot/batch: 05/2022
- Stability under test conditions: stable
- Storage condition of test material: dry conditions

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeding farm BioTest s.r.o., Konarovice, 281 25, Czech Republic,
- Weight at study initiation: 202-235 g
- Housing: animal room with monitoring conditions – one animal in one plastic cage
- Diet (e.g. ad libitum): ST 1 BERGMAN – complete pelleted diet ad libitum
- Water (e.g. ad libitum): drinking tap water ad libitum
- Acclimation period: 8 days
- Bedding: sterilized shavings of soft wood
- Randomisation: according to the internal rule, at the start of the study the weight variation of animals was minimal and did not exceed + 20 % of the mean weight
- Identification of animals: colour marks on tail, each cage was marked with the number of study, number of animal, sex, name and dose of the test
substance
- Health condition: certificate of good health condition – from breeding farm; no signs of diseases were observed at clinical check-in, during the acclimatisation period and before the start of study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): room temperature 22 +/- 3°C, permanently monitored
- Humidity (%): relative humidity 30 – 70 %, permanently monitored
- Photoperiod (hrs dark / hrs light): 12-hour light/12 hour dark

Administration / exposure

Type of coverage:
occlusive
Vehicle:
other: No vehicle, the substance was moistened with the smallest amount of water
Details on dermal exposure:
TEST SITE
- Area of exposure: 6x6 cm
- % coverage: 10%
- Type of wrap if used: covered by mull, plastic foil and held in contact by plaster (strapping)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): washing with water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): The substance was tested as such, moistened with the smallest amount of water. 2000 mg/kg

VEHICLE
no vehicle
Duration of exposure:
24 hours
Doses:
2000 mg/kg
No. of animals per sex per dose:
10 animals: 5 females+ 5 males
Control animals:
no
Details on study design:
- Duration of observation period following administration: 15 days
- Frequency of observations and weighing:
Observation: the first day: twice (30 minutes and 3 hours after application), the second day: twice (in the morning and in the afternoon) and daily thereafter for 14 days.
Observations included changes in skin and fur, eyes, visible mucous membranes, nutritive condition, autonomic and central nervous systems, psychic activity, somatomotor activity, reactions to stimuli, function of respiratory, circulatory, digestive and urogenital system. Also presence of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma was carefully observed.
Weighing: The animals were weighed at the start of the study (before application), at 8th day and in the end of experiment (15th day). Average body weight in-group was counted from individual body weights.
- Necropsy of survivors performed::
All test animals survived to the end of study were sacrificed on the 15th day by injection of veterinary preparation T61 (1 ml iv.) and gross necropsy was carried out. Nutritious status, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated.

Results and discussion

Preliminary study:
no
Effect levelsopen allclose all
Sex:
male
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
No mortality
Clinical signs:
No clinical signs of toxicity were observed during the study in all animals.
In one test animal the skin irritation symptoms (very slight erythema about large 0.5 cm2) of the test area were observed at 30 minutes after end application in scheduled time intervals. Three hours after end application erythema extinction fully. Change on the skin was reversible.
Body weight:
Weight increments in all animals were in physiological range.
Gross pathology:
No death of animals was observed during 14 days observation period.
Macroscopic changes were not diagnosed during pathological examination near all animals in both groups. In two test animals (1 male and 1 female) viscous content of intestine (diarrhoea) and in one female dilatation of uterus (demonstration of oestrus cycle) were found out.

Any other information on results incl. tables

Pathological examination  2000 mg/kg males 

Animal No.

         

Findings

1

Without pathologic changes

2

Without pathologic changes

3

Without pathologic changes

4

Without pathologic changes

5

Without pathologic changes, (viscous content of intestine (diarrhoea))

Pathological examination  2000 mg/kg females

 

Animal No.

         

Findings

1

Without pathologic changes

2

Without pathologic changes, (viscous content of intestine (diarrhoea))

3

Without pathologic changes

4

Without pathologic changes, (Uterus – dilatation, demonstration of oestrus cycle)

5

Without pathologic changes

 

Applicant's summary and conclusion

Interpretation of results:
practically nontoxic
Remarks:
Migrated information Criteria used for interpretation of results: EU
Executive summary:

The test substance was tested for the assessment of acute dermal toxicity using Wistar rats.

Testing was performed according to the Method B.3 Acute Toxicity (Dermal), Directive 92/69/EEC, published in OJ L 383A, 1992.

The study was performed as limit test: two groups of animals – 5 males and 5 females and the dose of 2000 mg/kg. The test substance was applied on the shaved skin of the test animals in delivered form (moistened with the smallest amount of water) for 24 hours.

The test animals were observed 14 days after application, afterwards were sacrificed and the necropsy for macroscopic examination of the organs was performed.

The test substance applied in dose 2000 mg/kg did not cause the death of animals. Clinical signs of toxicity were observed during the study in one animal (very slight erythema). Macroscopic changes not were diagnosed during pathological examination in all animals.

According to the results of study the value of LD50 dermal of the test substance for rats of both sexes is higher than 2000 mg/kg.