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Short-term toxicity to aquatic invertebrates

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Reference
Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
other: ISO TC147/SC5/WG2 "Water Quality, Determination of Acute Lethal Toxicity to Marine Copepods (Copepoda, Crustacea)" (Now ISO 14669)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
no
Details on sampling:
- Concentrations: 0 and 1000 mg/l nominal WAF loading rate (0, 10, 100 and 1000 mg/l nominal WAF loading rate in the range-finding study)
- Sampling method: The concentration, homogeneity and stability of the test material in the test preparations were not determined. Total Organic Carbon (TOC) analysis was performed on the test preparations at 0 and 48 hours. Duplicate samples were taken and stored at -20°C for further analysis if necessary.
- Sample storage conditions before analysis: Not reported
Vehicle:
no
Details on test solutions:
PREPARATION OF SYNTHETIC SEAWATER
Tropic Marin(R) Synthetic Sea Salt (750 g) was dispersed in a volume of 20 litres of water, giving a nominal salinity of 31.6 ppt. The pH was adjusted to 7.9 using 10 M HCl.

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 2.50 g of test material was added to the surface of 2.5 litres of synthetic seawater (prepared using Tropic Marin(R) Synthetic Sea Sailt). The synthetic seawater was then stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. Stirring was conducted for 16 hours. The mixture was then allowed to stand for 4 hours. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel to a depth of 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase or WAF was removed by mid-depth siphoning (the first 75-100 ml discarded) to give the 1000 mg/l loading rate WAF.
- Evidence of undissolved material: Microscopic inspection of the WAF showed no micro-dispersions or undissolved test material to be present.
Test organisms (species):
other aquatic crustacea: Acartia tonsa
Details on test organisms:
TEST ORGANISM
- Common name: Copepod
- Source: Guernsey Sea Farms, Vale, Guernsey, UK
- Age at study initiation: 12-14 days on day of supply
- Weight at study initiation: Not stated
- Length at study initiation: Not stated
- Feeding during test: Organisms were not fed during the test


ACCLIMATION
- Acclimation period: None reported
- Acclimation conditions: Prior to dispatch, organisms were held in sea water with a salinity of 33 ± 2 ppt at 20 ± 1°C
- Type and amount of food: Prior to dispatch, organisms were fed on a mixed culture of marine algae
Test type:
static
Water media type:
saltwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
Not reported
Hardness:
Not relevant
Test temperature:
Refer to Table 1
pH:
Refer to Table 1
Dissolved oxygen:
Refer to Table 1
Salinity:
31.6 ppt in the definitive test, 31.8 ppt in the range-finding study
Nominal and measured concentrations:
0 and 1000 mg/l nominal WAF loading rate
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Material: 50 ml glass jar containing 25 ml of test preparation, covered to reduce evaporation
- Aeration: Not reported
- Renewal rate of test solution: Test solution was not renewed
- No. of organisms per vessel: Five
- No. of vessels per concentration (replicates): Eight
- No. of vessels per control (replicates): Four
- Biomass loading rate: Not reported


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic seawater. Tropic Marin(R) Synthetic Sea Salt (750 g) was dispersed in a volume of 20 litres of water, giving a nominal salinity of 31.6 ppt. The pH was adjusted to 7.9 using 10 M HCl.
- Total organic carbon: Refer to Table 2
- Intervals of water quality measurement: 0 and 48 hours (pH, dissolved oxygen); 0, 24 and 48 hours (temperature)


OTHER TEST CONDITIONS
- Adjustment of pH: Not stated
- Photoperiod: 16 hour light, 8 hour darkness with 20 minute dawn and dusk transitions
- Light intensity: Not stated


EFFECT PARAMETERS MEASURED: Mortality or adverse reactions to exposure. Mortality was considered to have occurred if Acartia tonsa were unable to swim for approximately 10 seconds after gentle agitation


TEST CONCENTRATIONS
- Test concentrations: 0 and 1000 mg/l nominal WAF loading rate
- Range finding study
- Test concentrations: 0, 10, 100 and 1000 mg/l nominal WAF loading rate
- Results used to determine the conditions for the definitive study: No mortality occurred in the range-finding study
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Duration:
24 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
NOELR
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: WAF loading rate
Basis for effect:
mortality
Details on results:
No mortalities occured at a concentration of 1000 mg/l nominal WAF loading rate
Results with reference substance (positive control):
- Results with reference substance valid?: Yes
- Mortality: 35% mortality after 24 hours, 70% mortality after 48 hours. Refer to Table 3.
Reported statistics and error estimates:
Statistical analysis of the data was not warranted on the basis that no mortality occurred in this study.

Table 3: Cumulative mortality data in the positive control

 

Cumulative mortality (initial population 5 per replicate)

24

48

Replicate

1

2

3

4

Total

%

1

2

3

4

Total

%

Positive control (1 mg/l)

1

3

1

2

7

35

3

5

2

4

14

70

Conclusions:
The test substance was not toxic to the Copepod (Acartia tonsa) at a loading rate of 1000 mg/L nominal WAF
Executive summary:

In a 48-hour acute toxicity study, Copepod (Acartia tonsa) were exposed to Cardolite NC-603 (technical grade)at nominal concentrations of 0 and 1000 mg/L WAF under static conditions.  The 48-hour LL50was >1000 mg/L nominal WAF.

 

This toxicity study is classified as acceptable and satisfies the guideline requirement for a invertebrate acute toxicity study.

Results Synopsis

Test Type: Static

LL50:  >1000 mg/L nominal WAF

NOELR:  1000 mg/L nominal WAF

Description of key information

Key value for chemical safety assessment

Additional information

In a 48-hour acute toxicity study, Copepod (Acartia tonsa) were exposed to Cardolite NC-603 (Technical Grade) at nominal concentrations of 0 and 1000 mg/L WAF under static conditions. 


 


No mortalities occurred at a concentration of 1000 mg/l nominal WAF loading rate.


 


The 48-hour LL50 was >1000 mg/L nominal WAF.


 


The study is classified as Klimisch Code 2 (reliable with restriction) as it was performed to an internationally recognised method (ISO 14669).