Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 February - 3 August 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
(No chemical analysis was conducted. The test substance is an UVCB, and no chemical analysis is available for the whole compound. Therefore, in agreement with the sponsor, no chemical analysis was performed.)
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
yes
Remarks:
(No chemical analysis was conducted. The test substance is an UVCB, and no chemical analysis is available for the whole compound. Therefore, in agreement with the sponsor, no chemical analysis was performed.)
Principles of method if other than guideline:
A threshold approach according to OECD draft guidance document ("The Threshold Approach for Acute Fish Toxicity Testing", December 2009) was applied to avoid unnecessary use of fish. Therefore a limit-test with a concentration of nominal 9 mg test substance per L dilution water was performed. The selection of this concentration was based on the lowest EC50 value derived from the studies of "GREEN LIQUOR 1" on algae and daphnia.
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Sulfite liquors and Cooking liquors, green
EC Number:
268-612-2
EC Name:
Sulfite liquors and Cooking liquors, green
Cas Number:
68131-30-6
Molecular formula:
HNa3OS
IUPAC Name:
Aqueous alkaline solution of sodium salts produced by dissolving melt from incineration of spent liquor from sodium based pulping processes.
Details on test material:
- Name of test material (as cited in study report): Green liquor 1 (Sulphite process)
- Chemical name: Sulphite liquors and Cooking liquors, green.
- Molecular formula (if other than submission substance): UVCB
- Appearance: Yellowish liquid
- pH: over 11
- Solubility: In water: The substance is a water solution
- Impurities (identity and concentrations): UVCB
- Lot/batch No.: Not stated
- Expiry date: Not available.
- Stability under test conditions: No data available
- Storage condition of test material: Room temperature. Storage in the dark but may be used under light.

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test media were prepared at each point of time (start, 24, 48, and 72 hours) by diluting nominal 45 mg test substance in 5 litres of dilution water by manual homogenisation.
- Controls: dilution water (local tap water + deionised water, 1:1)
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): No.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Strain: not applicable
- Source: Tomy´s Zoo GmbH, Josef-Strebl-Gasse 6, 2345 Brunn am Gebirge
- Length at study initiation (length definition, mean, range and SD): 3.1 cm, SD: 0.3 cm (mean of control group at end of study)
- Weight at study initiation (mean and range, SD): 0.56 g, SD: 0.06 (mean of control group at end of study)
- Feeding during test: None

ACCLIMATION
- Acclimation period: Approx. 6 weeks
- Acclimation conditions (same as test or not): Same; Holding: glass vessel (60 cm x 30 cm x 30 cm), each filled with
ca. 54 L and covered with a transparent plastic lid. Water was purified by recirculation (power filter pump EHEIM/JBL) through filter wool.
Exposure: full glass vessels (29 cm x 20 cm x 24 cm), filled with
5 L of control and test medium and covered with transparent plastic lids.
- Type and amount of food: TetraMin, Tetra Werke, Ultrich Baensch GmbH, D-49304 Melle. feeding ad libitum
- Feeding frequency: Daily (Exposure: no feeding)
- Health during acclimation (any mortality observed): Mortality of less than 5 %.

Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Post exposure observation period:
None

Test conditions

Hardness:
210-230 mg CaCO3/L
Test temperature:
19 - 21 °C
pH:
7.9 to 8.3 in control medium, 7.8 to 8.4 in test medium.
Dissolved oxygen:
At least 77 % of saturation value at corresponding temperature.
Salinity:
Not applicable
Nominal and measured concentrations:
Nominal (mean actual): 9.2 mg/L (wwt) 1.6 mg/l (dw) (The dry solids content of the tested sample is 17.7% dry solids/82.3% water by weight (at 20 deg. C)).
Details on test conditions:
TEST SYSTEM
- Test vessel: full glass vessels, filled with 5 L control/ test medium and covered with transparent plastic lids.
- Material, size, headspace, fill volume: glass vessels, 29 cm x 20 cm x 24 cm
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): 1x/d
- No. of organisms per vessel: 7/vessel
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: max. 1 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: local tap water mixed with deionised water to achieve the appropriate water hardness required by the guideline. Mixing ratio: 1:1
- Culture medium different from test medium: No
- Intervals of water quality measurement: 1x/d, before and after media change

OTHER TEST CONDITIONS
- Adjustment of pH: No.
- Photoperiod: artificial light from 4 am to 8 pm

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : All animals were observed for alterations and behavioural changes at introduction in the test vessels and approximately 2, 4, 24, 48, 72 and 96 hours afterwards. At 24, 48, and 72 hours the fish were observed before and after the media change.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: According to the OECD draft guidance document ("The Threshold Approach for Acute Fish Toxicity Testing", December 2009) the lowest EC50 value of existing and reliable algae or acute invertebrate toxicity data is set as threshold concentration. An acute fish limit test is performed at the threshold to consider if fish are more or less sensitive than groups/species for which an EC50 is available.
- Range finding study: Two separate Seibersdorf Labor GmbH studies on algae (SL-LT-191/10) and daphnia
(SL-LT-221/10) for "GREEN LIQUOR 1" were performed.
- Results used to determine the conditions for the definitive study: The EC50/48h for daphnia is nominal 9 mg/L (wwt.) 1.6 mg/l (dw) and the EC50/72h for algae based on the average growth rates is nominal 1380 mg/L (wwt) 244 mg/l (dw.). Therefore a nominal threshold concentration of 9 mg/L(wwt.) 1.6 mg/l (dw.) was used for the acute toxicity study for fish.
Reference substance (positive control):
no

Results and discussion

Effect concentrationsopen allclose all
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LC50
Effect conc.:
> 9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
EC50
Effect conc.:
> 9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
> 9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
>= 9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
9.2 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
wet weight
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: None.
- Observations on body length and weight: Length: 3.1 cm (mean of control group), weight: 0.56 g (mean of control group)
- Other biological observations: No.
- Mortality of control: None.
- Other adverse effects control: No.
- Abnormal responses: None.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No.
- Effect concentrations exceeding solubility of substance in test medium: No.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
Not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
LC50, 96 h: > 9.2 mg/L wet weight, > 1.63 mg/L dry weight
LC100, 96 h: > 9.2mg/L wet weight, > 1.63 mg/L dry weight
LC0, 96 h: >= 9.2 mg/L wet weight, >= 1.63 mg/L dry weight
NOEC: 9.2 mg/L wet weight, 1.63 mg/L dry weight
Executive summary:

The acute toxicity of "GREEN LIQUOR 1" toZebrafish (Danio rerio) was determined for regulatory purposes.

The investigation was conducted according to the Regulation (EC) 440/2008: C.1. Acute Toxicity for Fish and OECD-Guideline 203, "Fish, acute toxicity test" (17thJuly 1992).

A threshold approach according toOECD draft guidance document ("The Threshold Approach for Acute Fish Toxicity Testing", December 2009) was applied to avoid unnecessary use of fish. Therefore alimit-test with a concentration of nominal 9 mg (wet weight basis, 82.3 % water) test substance per L dilution water was performed. The selection of this concentration was based on the lowest EC50value derived from the studies of"GREEN LIQUOR 1"on algae and daphnia.

7 Zebrafish(Danio rerio) were exposed in the test substance group and 7 in the control group. Their length was between 3.0 and 3.3 cm.

A nominal test substance concentration of 9.2 mg/L (wwt)/ 1.63 mg/L (dw.) was prepared by diluting the test substance in dilution water. (The dry solids content of the tested sample is 204 g/l at 20 deg. C (17.7% dry solids/82.3% water by weight)). Dilution water was also used as the negative control substance and for holding of the fish before the test.

The dilution water was local tap water which was mixed 1:1 with deionised water to achieve the appropriate water hardness as required by the guideline. Dilution water was also used for the negative control group.

The exposure of the Zebrafish was for 96 h in a semi-static test with renewal of test and control media after 24, 48 and 72 hours.

Results

LC50, 96 h: > 9.2 mg/L (wwt.), > 1.6 mg/L (dw.)

LC100, 96 h: > 9.2mg/L (wwt.), > 1.6 mg/L (dw.))

LC0, 96 h: >= 9.2 mg/L (wwt.), >= 1.6 mg/L (dw.)

NOEC: 9.2 mg/L (wwt.), 1.6 mg/L (dw.)