Ashes (residues), nonhazardous municipal solid waste

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

7.1.2008-25.1.2008

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 C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

The study was perfomed without the analytical determination of the test substance concentrations. For all evaluations and results nominal concentrations of the test substance were used.

GLP compliance:

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

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION OF SOLUTIONS OF TEST SUBSTANCE:

Since the test substance is poorly soluble in dilution water at concentrations needed for the test, the saturated solution of the test substance in dilution water was used as stock solution. 200 mg of the test substance was weighted into 2000 mL of dilution water for preliminary test and 1000 mg of the test substance was weighted into 10000 mL of dilution water for limit test. This mixture was stirred on a shaft stirrer throughout three days, ultrasonicated for 10 minutes and filtered through 0.45 micron filter. The concentrations of solutions used in the preliminary and limit test were obtained by dilution of the stock saturated solution with dilution water.

Test organisms

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Strain: Peters
- Source: certified supplier BioTest s.r.o., Konárovice (Czech Republic)
- Age at study initiation: 3-4 monts
- Length at study initiation : 2.0-3.0 cm
- Method of breeding: The fish were kept in all-glass aquaria in aerated drinking water
- Feeding during test: without feeding


ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: ad libitum "Tropica" - feed for aquarium fish, sometimes live daphnia
- Health during acclimation (any mortality observed): healthy animals

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

Preliminary test: Content of (Ca+Mg) ions 2.61 mmol.L-1
Limit test: Content of (Ca+Mg) ions 2.66 mmol.L-1

Test temperature:

22+/-2oC, with a variationof +/-1oC at each individual test

pH:

Preliminary test: pH of dilution water 7.9
Limit test: pH of dilution water 7.9

Dissolved oxygen:

Preliminary test: Oxygen content 7.6 mg.L-1 at temperature of 21oC
Limit test: Oxygen content 8.6 mg.L-1 at at temperature of 21oC

Nominal and measured concentrations:

Since the test substance is poorly soluble in dilution water it was not possible to achieve the concentration of 100 mg.L-1. For this reason a saturated solution of the test substance in dilution water was prepared for preliminary and limit test.

Preliminary test: nominal concentration range 1-100 mg.L-1 (100, 50, 10, 5, 1 mg.L-1)
Limit test: nominal concentration range 62-100 mg.L-1 (100, 91, 83, 75, 68, 62 mg.L-1)

Details on test conditions:

TEST SYSTEM
- Test vessel: Type: open
- Material, fill volume: glass beakers, fill volume 2 L
- Aeration: without aeration
- No. of organisms per vessel: Preliminary test 7. Limit test 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 1 g per liter


TEST MEDIUM / WATER PARAMETERS
Standard dilution water was prepared just before the test.
The dilution water should be of pH 7.8 +/- 0.2. It is saturated by oxygen by the aeration at temperature of 20 - 24°C. The content of sum Ca + Mg ions should have the value of 2.5 mmol.L-1.


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: natural variation of daily light and dark


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Fish mortality was observed each 24 hours. The pH, temperature and dissolved oxygen were measured at the same intervals.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.1
- Justification for using less concentrations than requested by guideline: Since the test substance is poorly soluble in dilution water it was not possible to achieve the concentration of 100 mg.L-1. For this reason a saturated solution of the test substance in dilution water was prepared for preliminary and limit test.
- Range finding study
- Test concentrations: 100, 91, 83, 75, 68, 62 mg.L-1 (nominal concentration)

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 5

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 6

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 7

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 8

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 9

Duration:

24 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 10

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 11

Duration:

72 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 12

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: not observed
- Observations on body length and weight: not observed
- Other biological observations: not observed
- Mortality of control: none
- Other adverse effects control: not observed
- Abnormal responses: not observed

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Mortality: yes
- LC50: LC50 (48h) 143 mg.L-1, LC50(96h) 137 mg.L-1

Any other information on results incl. tables

Sublethal observations / clinical signs:

Limit test

Table 4: Fish mortality – limit test

Nominal concentration	mg·L^-1	100	91	83	75	68	62	C
Saturated solution dosing	mL·L^-1	1000	910	830	750	680	620	C
Number of live fish	2 hours	10	10	10	10	10	10	10
Mortality %	2 hours	0	0	0	0	0	0	0
Number of live fish	24 hours	10	10	10	10	10	10	10
Mortality %	24 hours	0	0	0	0	0	0	0
Number of live fish	48 hours	10	10	10	10	10	10	10
Mortality %	48 hours	0	0	0	0	0	0	0
Number of live fish	72 hours	10	10	10	10	10	10	10
Mortality %	72 hours	0	0	0	0	0	0	0
Number of live fish	96 hours	10	10	10	10	10	10	10
Mortality %	96 hours	0	0	0	0	0	0	0

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Study evaluation
No mortality was observed in limit test. Therefore, exact values of LC50 could not be calculated and the values of LC are given in the form of a range. The NOEC values are identified by direct observation.
The nominal concentrations of the test substance were used for all evaluations and results.

Executive summary:

The test substance, Ashes (residues), was tested for acute toxicity on fish Poecilia reticulata.

The test was performed according to EU Method C.1., Acute toxicity for fish, Directive 92/69/EEC. Published in O.J. L 383A, 1992.

 

Since the test substance is poorly soluble in dilution water it was not possible to achieve the concentration of 100 mg·L^-1. For this reason a saturated solution of the test substance in dilution water was prepared for preliminary and limit test.

 

The preliminary test was performed in range of test substance nominal concentrations 1 – 100 mg·L^-1. Based on no toxicity of the test substance found in preliminary test, the limit test was performed. The limit test was performed in range of test substance nominal concentrations 62 – 100 mg·L^-1. The test was performed as static.

 

The study was performed without the analytical determination of the test substance concentrations. The nominal concentrations of the test substance were used for all evaluations and results.

 

Test results:

24h – LC₅₀>100 mg·L^-1(nominal concentration)

48h – LC₅₀>100 mg·L^-1(nominal concentration)

72h – LC₅₀>100 mg·L^-1(nominal concentration)

96h – LC₅₀>100 mg·L^-1(nominal concentration)

24h – NOEC=100 mg·L^-1(nominal concentration)

48h – NOEC=100 mg·L^-1(nominal concentration)

72h – NOEC=100 mg·L^-1(nominal concentration)

96h – NOEC=100 mg·L^-1(nominal concentration)

24h – LC₁₀₀>100 mg·L^-1(nominal concentration)

48h – LC₁₀₀>100 mg·L^-1(nominal concentration)

72h – LC₁₀₀>100 mg·L^-1(nominal concentration)

96h – LC₁₀₀>100 mg·L^-1(nominal concentration)

 

Classification of the test substance on the basis of hazard to aquatic environment was performed according to the Directive 93/21/EEC, Annex IV (article 5.2.1.).

 

According to the classification criteria given in quoted Directive the test substance, Ashes (residues), is not assigned to any of quoted toxicity categories.