Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15.2.2010-2.3.2010
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:
2010
Report Date:
2010

Materials and methods

Test guideline
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
Details on test material:
- Name of test material: Ashes (residues) – Biomass Combustion
- Physical state: solid
- Substance type: technical product
- Appearance: black powder

- Composition of test material, percentage of components:
SiO2 37.81 % (w/w), K2O 17.01 % (w/w), CaO (total) 11.92 % (w/w), P2O5 6.15 % (w/w), Al2O3 4.05 % (w/w), SO3 (sulphate) 3.47 % (w/w), MgO 2.25 % (w/w), Fe2O3 1.81 % (w/w), CaO (free) 1.40 % (w/w), CO2 1.29 % (w/w), Na2O 0.60 % (w/w), TiO2 0.34 % (w/w).
- Impurities (identity and concentrations):
Metals: As, Be, Cd, Co, Cr, Cu, Hg, Mo, Ni, Pb, Sb, Se, Tl, V, Zn sum <
0.1%

- Lot/batch No.: BMA/F/2009
- Expiration date of the lot/batch: 15 years / 12/2024
- Storage condition of test material: PE container, temperature bellow 40°C

Test animals

Species:
rat
Strain:
Wistar
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: breeding farm VELAZ, s.r.o., Koleč u Kladna, Czech Republic, RČH CZ 21760152
- Age at study initiation: 8-10 weeks
- Weight at study initiation: see Table 1
- Fasting period before study: cca 20 h
- Housing: animal room with monitoring conditions – 3 animals of one sex in one plastic breeding cage Velaz T4
- Diet (e.g. ad libitum): ST 1 BERGMAN – standard pelleted diet ad libitum, (producer: Ing.Mrkvička Miroslav - Výroba krmných směsí, Mlýn Kocanda, 252 42 Jesenice u Prahy)
- Water (e.g. ad libitum): drinking tap water ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law)
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C (permanently monitored)
- Humidity (%): 30-70 % (permanently monitored)
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours light / 12 hours dark

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
VEHICLE
- Olive oil; Lot/batch no.: 4683401
- Expiration.: 04/2011

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw

DOSAGE PREPARATION:
Immediately before application the test substance was weighed, mixed in vehicle (olive oil) and resulting suspension was administered to the stomach by tube. The single volume of administered suspension was 1ml/100 g of animal body weight.

CLASS METHOD:
- Rationale for the selection of the starting dose: according to methodology
Testing schedule (according to EU Method B.1 tris Annex 1D)
START: 2000 mg/kg – 3 females (Step No.1): no deaths ► 2000 mg/kg – 3 females (Step No. 2): no deaths ► END of study
Doses:
2000 mg/kg (bw)
No. of animals per sex per dose:
3 rats
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Mortality: daily
- Frequency of observations: daily, after aplication the animals were observed individually
the first day: twice (30 minutes and 3 hours after application)
the second day: twice (in the morning and in the afternoon), daily thereafter for 14 days
Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, and presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.

- Frequency of weighing:
before application, the 8th day and before euthanasia of animals (see Table 1)

- Necropsy of survivors performed: yes
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

Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
no death animals
Clinical signs:
no clinical signs of intoxication
Body weight:
see Table 1
Gross pathology:
without pathologic changes

Any other information on results incl. tables

Table 1: Test results

Dose (mg/kg) 

(Step No.)

Animal

No.

Body weight (g)

Body weight gain (g)

Before
application

8 days
p.a.

15 days
p.a.

0-8 days
p.a.

8-15 days
p.a.

2000

(1)

1

158.30

187.40

202.62

29.10

15.22

2

144.68

179.79

194.94

35.11

15.15

3

159.02

191.22

205.17

32.20

13.95

mean

154.00

186.14

200.91

32.14

14.77

SD

8.08

5.82

5.33

3.01

0.71

2000

(2)

4

163.75

199.26

218.03

35.51

18.77

5

153.00

177.99

190.60

24.99

12.61

6

155.68

191.12

205.87

35.44

14.75

mean

157.48

189.46

204.83

31.98

15.38

SD

5.60

10.73

13.74

6.05

3.12

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance toxicity was evaluated on the basis of mortality, clinical signs of intoxication, body weight increments during the observation period and necropsy findings at the end of study. The test substance administered at the dose of 2000 mg/kg caused no death of animals.
No clinical signs of intoxication were observed during study in all six animals.
No pathologic macroscopic changes were diagnosed during pathological examination.
According to the study results the value of LD50 of the test substance, Ashes (residues) – Biomass Combustion, (in female rats) is higher than 2000 mg/kg of body weight.
Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance,Ashes (residues) – Biomass Combustion,after a single oral administration to Wistar rats.

The testing was performed according to the Method B.1 tris: Acute Oral Toxicity - Acute Toxic Class Method, Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

Test substance wasadministered in a single dose as a solution in olive oil (vehicle), given orally via gavage to two groups of three female Wistar rats.

The dosing was performed sequentially in two groups of three females: group No. 1 - first step using the starting dose of 2000 mg/kg of body weight and group No.2 - second step using the same dose.

The test substance administered at the dose of 2000 mg/kg caused no death of animals.  

No clinical signs of intoxication were observed during study in all six animals.

No pathologic macroscopic changes were diagnosed during pathological examination.

According to the study results the value of LD50of the test substance for female rats is higher than 2000 mg/kg of body weight.