Registration Dossier

Administrative data

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23. 08. – 08. 09. 2010
Reliability:
1 (reliable without restriction)

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.3 (Acute Toxicity (Dermal))
Deviations:
yes
Remarks:
(See Overall remarks)
GLP compliance:
yes
Test type:
standard acute 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:
other: Wistar han
Sex:
male/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
- Weight at study initiation: see Table 1 and 2 in Any other information on results part
- Housing: animal room with monitoring conditions – 1 animal in one plastic cage
- 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)

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

Type of coverage:
semiocclusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: 6x6 cm
- % coverage: 10
- Type of wrap if used: covered by mull and held in contact by plaster

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

TEST MATERIAL
in delivered form
amount was weighed out - 2000 mg/kg bw


Duration of exposure:
24 hours
Doses:
2000 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: mortality (daily), clinical signs (daily), body weight ( before application, 8th and 15th day of study)
- Necropsy of survivors performed: yes (15th day of study)

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
daily
Clinical signs:
After application the animals were observed individually – at the first day: twice (30 minutes and 3 hours after application), at 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.
Body weight:
see Table 1 and 2 in Any other information on results part
Gross pathology:
All test animals surviving to the end of study were sacrificed on the 15th day by prolonged ethereal narcosis, and gross necropsy was carried out. Nutritional state, body surface, body foramina, thoracic, abdominal and cranial cavity were evaluated.

Any other information on results incl. tables

Weight of animals

In males, the weight increments were adequate to species and age of animals in the experiment (see table No.1).

Table No. 1: Individual body weight of animals – 2000 mg/kg – males

Animal No.

Before application

8th day

15th day

Body weight gain (g)

day 0-8
p.a.

day 8-15
p.a.

1 (pre-test)

248.29

280.39

312.23

32.10

31.84

2

250.62

292.41

327.87

41.79

35.46

3

245.44

268.27

301.74

22.83

33.47

4

252.82

276.38

306.94

23.56

30.56

5

249.29

281.01

317.21

31.72

36.20

Average

249.29

279.69

313.20

30.42

33.51

In females, the weight increments were adequate to species and age of animals in the experiment (see table No.2). Only slight decreased body weight and body weight increment in female No.4 and No.5 was recorded.

Table No. 2: Individual body weight of animals – 2000 mg/kg – females

Animal No.

Before application

8th day

15th day

Body weight gain (g)

day 0-8
p.a.

day 8-15
p.a.

1 (pre-test)

195.92

202.46

207.75

6.54

5.29

2

208.09

212.21

224.40

4.12

12.19

3

193.16

200.45

218.26

7.29

17.81

4

209.34

200.20

206.57

-9.14

6.37

5

198.22

196.86

209.27

-1.36

12.41

Average

200.95

202.44

213.25

1.49

10.81

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
According to the results of the dermal toxicity study, the value of LD50 (dermal) of Ashes (residues) – Biomass Combustion, for rats is higher than 2000 mg/kg of body weight.
Executive summary:

The test substance, Ashes (residues) – Biomass Combustion, was tested for acute dermal toxicity using Wistar rats.

Testing was performed according to the Method B.3 Acute Toxicity (Dermal), Council Regulation (EC) No.440/2008, published in O.J. L 142, 2008.

The study was performed as limit test: two groups of animals – 5 males and 5 females and the dose of 2000 mg/kg of body weight. The pre-test was performed with 1 male and 1 female from each group. After clinical observation of these pilot animals for 2 days after test substance application, the other animals of the group were dosed.

The test substance was applied on the shaved skin of the test animals in delivered form for 24 hours.

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

The test substance applied at the dose of 2000 mg/kg of body weight did not cause death of animals. No clinical signs of intoxication were observed in all animals. No macroscopic change was diagnosed during pathological examination in all animals.

According to the results of study, the value of LD50 (dermal) of the test substance, Ashes (residues) – Biomass Combustion, for rats of both sexes is higher than 2000 mg/kg of body weight.