Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2014-03-31 to 2014-04-08
Reliability:
1 (reliable without restriction)
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: The effects of the Water Accommodated Fraction (WAF) were tested in a limit test. The loading level was based on the results of a preliminary range finding test.
- Sampling method:
The analytical evaluation of the control, pH-control and the limit loading level of celitement was carried out via spectrophotometric analysis of silicon from freshly prepared test media at start of the exposure (0 h) and from corresponding 96 h aged test media at the end of the exposure.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: One Water Accommodated Fraction (WAF) was prepared with a nominal loading of 100 mg/L. The test item was applied onto a glass slide. The glass slide with the test item was inserted in a brown glass flask with an appropriate amount of dilution water. This dispersion was shaken for 24 hours with 20 rpm at room temperature. After a separation phase of 1 hour, the aqueous phase or WAF was removed by siphoning (from the approximate centre of the glass flask) to give the 100 mg/L loading rate WAF.


- Eluate: Dilution water
- Differential loading: Limit loading level of 100 mg/L
- Controls: 7 fish in dilution water (without test item) were tested under the same test conditions as the test replicates.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish
- Source: All fish used in the test were gained at DR.NOACK-LABORATORIEN from a single brood stock. (supplier: Umweltbundesamt, Schichauweg 58, D-12307 Berlin)
- Length at study initiation (length definition, mean, range and SD): Average body length: 2.71 cm
- Weight at study initiation (mean and range, SD): Average body weight: 0.39 g
- Method of breeding:Holding was performed at the test facility at 23 +/- 2 °C and diffuse light (0.1 - 10 µmol photons • m-2 • s-1, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value. No disease treatments were administered throughout holding and testing. Food was provided 3 times per week. The amount of food was 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started.
- Feeding during test: No feeding during test



ACCLIMATION
- Acclimation period: Zebrafish with at least 12 days of acclimatisation and mortality < 5 % within these days before the study starts were used in the test.
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: 4 % of the fish body weight per feeding day. The test fish were not fed 24 h before the test started. Food: Sera Vipan; SERA GMBH, D-52518 Heinsberg
- Feeding frequency: 3 times per week.
- Health during acclimation (any mortality observed): No mortality observed, no disease treatments were administered throughout holding and testing.


Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
Total Hardness at day 0: 49 mg/L
Test temperature:
Please refer to "Any other information on materials and methods"
pH:
Please refer to "Any other information on materials and methods"
Dissolved oxygen:
Please refer to "Any other information on materials and methods"
Salinity:
Not measured, freshwater
Nominal and measured concentrations:
Please refer to "Any other information on materials and methods incl. tables"
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass aquaria
- Type (delete if not applicable): open, loosely covered by glass tops
- Material, size, headspace, fill volume: glass, 3.1 L test volume per vessel
- Aeration: Gentle aeration
- Renewal rate of test solution (frequency/flow rate): None, static design
- Control: 7 fish in dilution water (without test item) were tested under the same test conditions as the test replicates
- pH-Control: One replicate of the limit loading level, neutralized with 1M NaOH and HCl to pH 7.0 ± 0.2, was tested under the same test conditions as the test item replicate.
- Reference item: No reference item is recommended for this test according to the guideline.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control and pH-control (replicates): 1
- Biomass loading rate: less than 1 g fish per L test solution


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water of local origin was used for holding and testing. The water was filtered on activated charcoal and aerated for at least 24 h to remove possible chlorine.
Nominal water parameters:
Total hardness: 10 - 250 mg CaCO3/L
pH-value: 6.0 - 8.5
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH-value, temperature and oxygen saturation were measured in all vessels at the beginning of the test and every 24 h. Total hardness of the water was determined at the beginning out of the test control. During the test the water temperature was recorded continuously with a data logger (once per hour).


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: natural
- Light intensity: 0.1 - 10 µmol photons x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fish were considered dead if there was no visible movement (e.g. gill covers movement) and if touching of the caudal peduncle produced no reaction. Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.). Observations were made after 24, 48, 72 and 96 h.


TEST CONCENTRATIONS
- Range finding study:
Cumulative Mortality [%] in the Preliminary Test
(n = 5)
Nominal test item loading level
[mg/L] Cumulative mortality [%] after different exposure periods [hours]
24 48 72 96
100 0 0 0 0
10 0 0 0 0
Control 0 0 0 0





Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LL0
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: Not observed
- Mortality of control: 0%
- Other adverse effects control: Not observed
- Abnormal responses: Not observed
- Any observations (e.g. precipitation): The test solution was turbid at test start and showed precipitation and sedimentation after 24 hours until the end of exposure.
Results with reference substance (positive control):
No reference substance tested
Reported statistics and error estimates:
The concentrations leading to 0 and 100 % mortality (LC0 and LC100) after 96 h were determined directly from the test results. LC10,20,50-values had not to be calculated in a limit test.

Observations in the Definitive Test

Nominal test item
loading level
[mg/L]

Obser-vations *



Effects on behaviour
after different exposure periods [hours]

24

48

72

96

100

(1)

7/7

7/7

7/7

7/7

100
(pH-control)

(1)

7/7

7/7

7/7

7/7

Control

(1)

7/7

7/7

7/7

7/7

Cumulative Mortality [%] in the Definitive Test

Nominal test item
loading level
[mg/L]

Cumulative mortality [%]
after different exposure periods [hours]

24

48

72

96

100

0

0

0

0

100
(pH-control)

0

0

0

0

Control

0

0

0

0

Validity criteria fulfilled:
yes
Conclusions:
In this study, celitement was found to cause no lethal effects to Zebrafish after 96 hours at a nominal test loading of 100 mg/L.
The LL50 after 96 hours is > 100 mg/L. All effect levels are given based on the nominal test item loading of the test item celitement.


Executive summary:

The acute toxicity of the test item celitement (batch no.: Not specified) to fish (zebrafish) was determined according to OECD-Guideline for Testing of Chemicals No. 203 (1992) from2014-03-31 to 2014-04-08, with the definitive exposure phase from2014-04-01 to 2014-04-05 at Dr.U.Noack-Laboratorien, 31157 Sarstedt, Germany.

A static limit test was performed with the Water Accommodated Fraction* (WAF) of 100 mg/L of celitement. One additional replicate of the limit loading level was prepared and neutralized with 1M NaOH and HCl to serve as pH-control. Duration of the test was 96 hours. 7 test organisms were exposed to the limit loading, the pH-control and the control. Water quality parameters pH-value, temperature and oxygen-saturation, measured after 0, 24, 48, 72 and 96 h were determined to be within the acceptable limits.

The concentrations of the test itemcelitementwere analytically evaluated via photometric analysis of silicon at the start of the exposure (0 h) and at the end of the exposure (96 h) of the limit loading level, the pH-control and the control.
The measured silicon concentration at the start of the exposure (0 h) was 7.95 mg/L in the limit loading level and 8.05 mg/L in the pH-control. At the end of the exposure the measured silicon concentrations gave recoveries of 95 and 99 % of the initially measured values of the limit loading and the pH-control indicating the stability of the test item in the WAF during the exposure phase

All effect levels are given based on the nominal loading level of the test item.

    

LL-Values (0 – 96 hours)

     Based on nominal test item loading [mg/L].

Test duration
[hours]

[mg/L]

LL10

LL20

LL50

24

> 100

> 100

> 100

48

> 100

> 100

> 100

72

> 100

> 100

> 100

96

> 100

> 100

> 100

LL0=

Highest test item loading with 0 % mortality after
96 hours

100

LC100=

Lowest test item loading with 100 % mortality after 96 hours

> 100

*)Water Accommodated Fraction (WAF): The aqueous medium containing only the fraction of a multi-component

  test item that is dissolved and / or present as a stable dispersion/emulsion under test conditions in the test

  medium, acc. to OECD Guidance Document, No. 23.

Description of key information

No acute (short term) toxicity to fish below a concentration of 100 mg/L, due to a Klimisch 1 study result.

Key value for chemical safety assessment

LC50 for freshwater fish:
100 mg/L

Additional information

Celitement did not cause any effects to zebrafish after 96 h when tested with a nominal test item concentration of 100 mg/L. Therefore the NOEC (0 – 96 h) is 100 mg/L and LC0 = 100 mg/L, LC50 > 100 mg/L, LC100 > 100 mg/L.