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Ecotoxicological information

Long-term toxicity to fish

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Administrative data

fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Basic data given: comparable to guidelines/standards, with analytical. Not enough details to warrant Klimisch 1 rating.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference Type:
study report

Materials and methods

Test guideline
equivalent or similar to
OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)
GLP compliance:
the study pre-dates GLPs

Test material

Details on test material:
- Name of test material: Two LAS test materials were used in this study:
1. 14C-radiolabeled C12LAS (pure C12 chainlength) (GTS72922), and
2. Unlabelled C11.6 LAS (comprised of C10,C11,C12,C13 chainlengths) (GTS72923)
- Substance type: Pure active substance
- Physical state: Not reported
- Specific activity: 831 µCi/mmole
- Locations of the label: uniformly labeled with 14 C in the benzene ring
- Stability under test conditions: Not reported
- Storage condition of test material: Not reported

Sampling and analysis

Analytical monitoring:
Details on sampling:
Please see the section below. The study report does not include details about sampling.

Test solutions

Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION: Preparation of test solutions is not reported. The test concentrations were delivered to the duplicate test vessels by a modified Mount and Brungs 1L proportional diluter.

Test organisms

Test organisms (species):
Lepomis macrochirus
Details on test organisms:
- Common name: Bluegill fish
- Strain: Not reported
- Source: Not reported
- Length at study initiation: 2.5-5.0 cm
- Age at study initiation: Juvenile
- Weight at study initiation: Not reported
- Method of breeding: Not applicable

- Food type: Frozen brine shrimp
- Amount: Fed at approx. 2% of total body weight/day
- Frequency: Daily

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
28 d
Post exposure observation period:

Test conditions

120 mg/L CaCO3
Test temperature:
21 ± 2 °C
8.1 ± 0.2
Dissolved oxygen:
Mean Dissolved Oxygen Concentration (DOC): 7.8 mg/L (ranged, 7.0-9.0)
Nominal and measured concentrations:
Nominal test concentrations were: 0, 0.5, 1.0, 2.0, and 4.0 mg/L. MBAS and radiometric counts confirmed the expected nominal concentrations. Two figures in the study report show the confirmation of nominal test concentrations, but details not provided.
Details on test conditions:
- Test vessel: Aquaria
- Type: Open
- Material: 19 L glass
- Aeration: No (test was flow-through)
- Type of flow-through: Proportional diluter
- Renewal rate of test solution: 8 replacement volumes/d
- No. of organisms per vessel: Five
- No. of vessels per concentration (replicates): Two
- No. of vessels per negative control (replicates): Two
- No. of vessels per vehicle control (replicates): Not applicable
- Biomass loading rate: Not reported. There were ¿5 pre-weighed juvenile bluegills per each aquarium,¿ according to the study report. Loading rate was therefore 1 juvenile bluegill/4 L test solution.

- Source/preparation of dilution water: The dilution water was carbon and reverse-osmosis filtered well water of 120 mg/L CaCO3 hardness.
- Total organic carbon: Not reported
- Particulate matter: Not reported
- Metals (mg/L): Zinc: <0.001; Lead: <0.01; Iron: <0.05; Copper: <0.001,
- Nitrate (mg/L): <0.05
- Pesticides: Not reported
- Chlorine: Not reported
- Alkalinity: Not reported
- Ca/mg ratio: Not reported
- Conductivity: Not reported
- Salinity: Not applicable
- Culture medium different from test medium: Not reported
- Intervals of water quality measurement: Twice weekly
- Other:
- Temperature was monitored continuously with a recording thermograph

- Adjustment of pH: Not reported
- Photoperiod: Not reported
- Light intensity: Not reported

EFFECT PARAMETERS MEASURED: (1) The growth of juvenile bluegills was determined from 10 individuals at each treatment concentration measured and weighed (wet weight) before and after the exposure. (2) Amount of 14C-LAS taken up by the fish was measured by combustion analysis.

NEGATIVE CONTROL PERFORMED: Yes (received only filtered well water)

- Sampling frequency: Only on Day 28, as sampling of test species were not carried out during exposure period, to avoid disturbance of the developing fishes. On Day 28, all 10 fish per test concentration were recovered.
- Sampling method: For the amount of 14C-LAS taken up by the fish, triplicate samples of the test species were taken at 28 d sampling period, rinsed, dried at 30°C for 24 h, and weighed in tared plancets prior to combustion analyses for radiometric counting of 14C-LAS content
- Sample storage conditions before analysis: For growth rate, fish were weighed immediately after conclusion of test. For 14C-LAS analysis, samples were dried at 30°C for 24 h before analysis

Reference substance (positive control):

Results and discussion

Effect concentrations
Key result
28 d
Dose descriptor:
Effect conc.:
1 mg/L
Nominal / measured:
Conc. based on:
act. ingr.
Basis for effect:
growth rate
Remarks on result:
other: Nominal concentration confirmed by analytical measurement
Details on results:
- Fish weights (individual and mean values) on day 0, 14 (if measured) and 28: Not reported
- Tank-average or pseudo-specific growth rates for periods 0-28 or 0-14 and 0-28 (if possible): Not reported
- Type of and number with morphological abnormalities: Not reported
- Type of and number with behavioural abnormalities: Not reported
- Other biological observations: Fish growth was reduced at both 2.0 and 4.0 mg/L test concentrations with no effect at 1.0 mg/L. At the end of the exposure period, test species had a biomass of 44 and 10.5 g/m2 at test concentration of 1 and 2 mg/L respectively. Control fish production was 42.5 g/m2
- Bioconcentration Data: The BCF value was found to be 252 based on a wet weight basis
- Effect concentrations exceeding solubility of substance in test medium: No
- Incidents in the course of the test which might have influenced the results: Not reported
Results with reference substance (positive control):
Positive control was not included in the study
Reported statistics and error estimates:
Not reported

Applicant's summary and conclusion

Validity criteria fulfilled:
The 28 d NOEC of C12 LAS to juvenile Bluegills (Lepomis macrochirus) was 1 mg/L, based on growth rate in a flow-through model ecosystem study. Analytical measurements confirmed the nominal test concentrations.
Executive summary:

A long term toxicity test to fish (juvenile Lepomis macrochirus) was conducted on C12LAS (linear alkylbenzene sulfonate),following the OECD 204 guideline. The growth of juvenile bluegills was determined after 28 days in a flow-through model ecosystem study.  The nominal test concentrations were 0, 0.5, 1.0, 2.0, and 4.0 mg/L, confirmed by analytical measurements.

The growth of juvenile bluegills (wet weight after 28 d exposure) was not affected at 0.5 and 1.0 mg LAS/L, but was reduced at 2.0 and 4.0 mg/L.  At the end of the exposure period, fish at 1.0 mg/L LAS had a biomass of 44 gm/m2 compared to 10.5 gm/m2 for the 2.0 mg/L concentration, and 42.5 gm/m2 for the controls.

The No observed effect concentration (NOEC) was 1.0 mg/L, based on growth rate.

This Chronic toxicity test is classified as acceptable, and satisfies the guideline requirements for the OECD 204.