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Short-term toxicity to fish

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short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study meets the criteria for Klimisch code 1 because it followed an internationally recognised method (OECD Guideline 203) and was carried out to Good Laboratory Practice.
according to
OECD Guideline 203 (Fish, Acute Toxicity Test)
None reported
GLP compliance:
Analytical monitoring:
Details on sampling:
Total organic carbon (TOC) measurements of each freshly prepared test solution and control and after 24-hours on test just before daily
renewal with fresh test solution.
Details on test solutions:
Individual water accommodated fractions (WAFs) were prepared for each daily renewal of the 10,000 mg/L test level.
A measured weight of test material was added to a measured volume of dilution water (15 L) in a glass vessel and stirred for 16 to 24 hour.
Stirring was accomplished using a Telfon coated magnetic stir bar.
Following the mixing period, the test solution were allowed to stand for 2 hours before the water phase was removed.
The water phase was siphoned from the middle of the vessels and was used for the toxicity test.

Dilution water was natural seawater collected from Cape Cod Canal, Bourne, Massachusetts. Water was filtered through a 0.5 micron polypropylene core filter and then activated carbon. Water was stored under constant aeration for 1-4 days prior to use.

No undissolved test material was reported on the surface of the test vessels during the entire toxicity test.
Test organisms (species):
Cyprinodon variegatus
Details on test organisms:
- Common name: Sheepshead minnow
- Source: Commercial supplier in New Hampshire
- Age at study initiation: 10 to 15 days old
- Length at study initiation: Average 11 mm, range 10-13 mm, n=30
- Weight at study initiation: Average 0.028 g, range 0.02-0.05 g, n=30
- Feeding during test: The fish were not fed during the test
- Loading biomass: 0.056 g biomass/L

- Acclimation period: Fish were held for a minimum of 6 days prior to testing
- Pretreatment: None
Test type:
other: Static renewal
Water media type:
Limit test:
Total exposure duration:
96 h
Post exposure observation period:
Not reported
Not relevant
Test temperature:
22 to 23°C
7.9 to 8.1
Dissolved oxygen:
5.5 mg/L to above 100% saturation (7.2 mg/L)
32 to 34 ppt
Nominal and measured concentrations:
Mean measured TOC levels in the control and 10,000 mg/L WAF test level were 3.0 mg/L (range 1.2 to 4.4) and 6.4 mg/L (range 5.0 to 7.9),
Details on test conditions:
- Test vessel: 5 L vessel
- Renewal rate of test solution (frequency/flow rate): About 90% of the test solution in each vessel renewed daily after 24, 48 and 72 hours.
- No. of organisms per vessel: Ten
- No. of vessels per concentration (replicates): Two

- Source/preparation of dilution water: Natural seawater collected from Cape Cod Canal, Bourne, Massachusetts

EFFECT PARAMETERS MEASURED: Mortality and signs of toxicity

- Test concentrations: Control and 10,000 mg/L WAF loading rate
Reference substance (positive control):
Sodium laurl sulfate (SLS).
Key result
96 h
Dose descriptor:
Effect conc.:
> 10 000 mg/L
Nominal / measured:
Conc. based on:
other: WAF loading rate
Basis for effect:
Key result
96 h
Dose descriptor:
Effect conc.:
> 10 000 mg/L
Nominal / measured:
Conc. based on:
other: WAF loading rate
Basis for effect:
Details on results:
Control mortality did not exceed the test validity criterion (i.e. <10% mortality in controls).

No mortality or signs of toxicity were noted in either the control or the 10,000 mg/L WAF treatment throughout the test.
Results with reference substance (positive control):
96 hour LC50 was 1.2 mg/L
Reported statistics and error estimates:
Statistical analysis of the data was not warranted on the basis that no mortality occurred in this study.
Validity criteria fulfilled:
The test substance was not toxic to the Sheepshead minnow (Cyprinidon variegatus) at a loading of 10000 mg/L nominal WAF
Executive summary:

In a 96-hour acute toxicity study, Sheepshead minnow (Cyprinidon variegatus) were exposed to benzenesulphonic acid C20-24 alkyl calcium salt at nominal concentrations of 0 and 10,000 mg/L WAF under static renewal conditions.  The 96-hour LL50was >10,000 mg/L nominal WAF. No signs of toxicity were noted.


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


Results Synopsis


Test organism size/age (mean wet weight or length): 10 to 15 days old, average length 11 mm average weight 0.028 g

Test Type: Static renewal


LL50:  >10,000 mg/L nominal WAF

LL0:  10,000 mg/L nominal WAF

Endpoint(s) Effected: None

Description of key information

The toxcity of the substance has been measured experimentally with freshwater fish.  Due to the extremely low water solubility all studies have been conducted using Water Accomodated Fractions and completed as limit tests.  

Key value for chemical safety assessment

LC50 for freshwater fish:
1 000 mg/L
LC50 for marine water fish:
10 000 mg/L

Additional information

The LC50 in freshwater fish is considered to be >1000 mg/l and >10000 mg/l with marine species. The substance is, therefore, considered not toxic to freshwater or marine fish species. Nevertheless, adequate controls will be in place during production and use to ensure that the substance is not released to the environment.