Ashes (residues), nonhazardous municipal solid waste

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7.1.2008-22.1.2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Report date:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

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

Test material information

Constituent 1

Reference substance name:

Ashes (residues)

EC Number:

268-627-4

EC Name:

Ashes (residues)

Cas Number:

68131-74-8

Test material form:

solid: bulk

Details on test material:

- Name of test material (as cited in study report): Ashes (residues)
- Substance type: technical product
- Physical state: solid
- Main components: SiO2 (42.12%), Al2O3 (32.16%), Fe2O3 (10.88%), TiO2, MnO, MgO, CaO, Na2O, K2O, SO3 - constituents with content less than 10%
- Expiration date of the lot/batch: unlimited
- Appearance: greyish black powder

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeding farm BioTest s.r.o., Konárovice, 281 25, CZ, RČH CZ 21760152
- Age at study initiation: 8-10 weeks at the time application
- Weight at study initiation: 164-167 g
- Fasting period before study: About twenty hours before oral administration the animals were not fed, water was given ad libitum.
- Housing: Animal room with monitoring conditions – 3 animals of one sex in one plastic breeding cage Velaz T4.
- Diet: ST 1 BERGMAN – standard pelleted diet ad libitum, (producer: Mill Kocanda, Jesenice u Prahy)
- Water: Drinking tap water ad libitum (quality corresponding to Regulation No. 252/2004 Czech Coll. of Law)
- Acclimation period: 6 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 for each sex.
- Identification of animals: Colour marks 1 - 3 on tail of animals, each cage was marked with the number of study, sex 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): 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-hour light/12 hour dark

STUDY TIME SCHEDULE
Animal supply: 02. 01. 2008
Experimental part of study: 07. 01. – 22. 01. 2008

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5% Methylcellulose in water

Details on oral exposure:

VEHICLE
- 0.5% methyl cellulose in water
- Justification for choice of vehicle: It is not toxic. Is it possible to prepared homogeneous suspension with test substance.
- Lot/batch no.: DT157078

MAXIMUM DOSE VOLUME APPLIED: 2000 mg/kg bw

DOSAGE PREPARATION:
Immediately before application the test substance was weighed, mixed in vehicle (0.5% methyl cellulose in water) 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 the methology
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:

6 females (Step No.1: 3 females, Step No.2: 3 females)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of weighing: Animals were weighed before application, on the 8th day of study and at day 15, before euthanasia of animals.
- Frequency of observation: After application 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) and daily thereafter for 14 days.
- Clinical examination: Observations included changes in skin and fur, eyes, visible mucous membranes, behaviour of animals, somatomotor activity, reactions to stimuli, presence of lacrimation, salivation and discharge from nostrils, function of respiratory, digestive and urogenital system.
- Pathological examination: 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.
- Body weight: Body weight increments were calculated from body weight at the start of the study and in the end of the study.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

No animals died during the study.

Clinical signs:

other: No clinical sings of intoxication were detected at 30 minutes and at 3 hours after application. In the next morning no clinical signs of intoxication were detected in all females. See Tables No. 2 and No. 3

Gross pathology:

All animals were without pathologic changes.

Any other information on results incl. tables

Table No. 2: Clinical observation – 2000 mg/kg (Step No.1)

Animal  No.	Death after application	Observed changes
1	No	30 minutes: no clinical signs of intoxication  3 hours:no clinical signs of intoxication 2^nd– 14^thday: no clinical signs of intoxication
2	No	30 minutes: no clinical signs of intoxication  3 hours: no clinical signs of intoxication 2^nd– 14^thday: no clinical signs of intoxication
3	No	30 minutes: no clinical signs of intoxication 3 hours: no clinical signs of intoxication 2^nd– 14^thday: no clinical signs of intoxication

Table No. 3: Clinical observation – 2000 mg/kg (Step No.2)

Animal  No.	Death after application	Observed changes
4	No	30 minutes: no clinical signs of intoxication  3 hours: no clinical signs of intoxication 2^nd– 14^thday: no clinical signs of intoxication
5	No	30 minutes: no clinical signs of intoxication  3 hours: no clinical signs of intoxication 2^nd– 14^thday: no clinical signs of intoxication
6	No	30 minutesno clinical signs of intoxication  3 hours: no clinical signs of intoxication 2^nd– 14^thday: no clinical signs of intoxication

Applicant's summary and conclusion

Interpretation of results:

practically nontoxic

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 in all animal. No pathologic macroscopic changes were diagnosed during pathological examination in all animals.

Executive summary:

The test substance was administered in a single dose as suspense in vehicle (0.5% methyl cellulose in water), 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 and group No.2 - second step using the starting dose of 2000 mg/kg body weight.The test substance administered at the dose of 2000 mg/kg causedno death of animals. No clinical signs of intoxication were observed in all six females. Macroscopic changes were not diagnosed during pathological examination in all females in both groups.

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