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Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

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Administrative data

Link to relevant study record(s)

Reference
Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
02/12/1993-04/23/1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study according to Official Jounal of the European Communities No. L133 Algal inhibition test.
Qualifier:
according to guideline
Guideline:
OECD Guideline 201 (Alga, Growth Inhibition Test)
Version / remarks:
1984
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Concentrations: 10, 3, 1, 0.4, 0.1 control mg/l
- Sample storage conditions before analysis: all samples were analyzed on the day taken
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test material and dilution water were stirred in sealed vessels for 24 hrs. The mixture was then allowed to settle for 1 hr. The aqueous phase was then drawn off and used as the test medium.
Test organisms (species):
Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)
Details on test organisms:
TEST ORGANISM
- Strain: axenic strain
- Source (laboratory, culture collection): American Type Culture Collection, Maryland, Ohio, USA
- Age of inoculum (at test initiation): in exponential growth phase
- Method of cultivation: 1.5% mass/volume agar is added to liquid medium and autoclaved. After cooling, the medium is added to 9 cm diameter sterile petri dishes. Cultures are streaked onto the plates and maintained at 18-22°C under continuous illumination for up to 2 weeks. Liquid medium cultures are made by tranferring cells on a sterile loop from the agar cultures. 100 ml batch cultures are grown in 250 ml Erlenmeyer flasks for up to 4 days maintained at 22-26°C under constant illumination of ~3000 lux in an incubator.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
72 h
Hardness:
Not determined
Test temperature:
21.9-24.3°C
pH:
7.2-10.3
pH exceeded preferred range in some test vessels. The flasks with good algal growth showed a large increase of pH (up to +3 pH units). Since they showed good growth, the high pH does not invalidate the test.
Dissolved oxygen:
Not determined
Salinity:
freshwater
Nominal and measured concentrations:
Nominal: 10, 3, 1, 0.4, 0.1, control mg/l
Measured: 9.4, 2.8, 1.3, 1.4, <1.0, <0.5 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed - sealed with glass stoppers
- Material, size, headspace, fill volume: 300 ml Erlenmeyer flask completely filled
- Aeration: None
- Renewal rate of test solution (frequency/flow rate): static
- Initial cells density: 5000 cells/ml
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6


GROWTH MEDIUM
- Standard medium used: Medium as described in EPA-600/9-78-01B except 105 µg boric acid, and 50 mg/l sodium bicarbonate. Prior to adding sodium bicarbonate, the medium was autoclaved at 1.0 kg/cm² for 15 min. After cooling, the sodium bircarbonate was added.
- Detailed composition if non-standard medium was used: algal nutrient medium augmented with sodium carbonate


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Millipore filtered water.

OTHER TEST CONDITIONS
- Light intensity and quality: ~3000 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: counted with Coulter Multisizer
Reference substance (positive control):
no
Key result
Duration:
72 h
Dose descriptor:
other: ErL50
Effect conc.:
2.9 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate
Remarks on result:
other: 2.6-3.3 mg/l
Key result
Duration:
72 h
Dose descriptor:
other: EbL50
Effect conc.:
2.6 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
biomass
Remarks on result:
other: 2.3-3.0 mg/l
Key result
Duration:
72 h
Dose descriptor:
NOELR
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: biomass and growth rate

 Summary of In-Life observations - % Inhibition                  

Loading Rate (mg/L)  Control   0.1 0.4 1 3 10
Based on Growth Rate 72 hrs  n/a 0  0  0  55  100
Based on Biomass 72 hrs   n/a  0  0  0  78  99
Validity criteria fulfilled:
yes
Conclusions:
The 72-hr ErL50 for R. subcapitata is 2.9 mg/l (WAF). The 72-hr EbL50 is 2.6 mg/l (WAF).
Executive summary:

This study was done to determine the acute toxicity of Hydrocarbons, C9, aromatics to the algae R. subcapitata. R. Subcapitata was exposed to 0, 0.1, 0.4, 1, 3, or 10 mg/l (WAF) of the test substance for 72 hrs. The mean cell concentration was measured at 0, 24, 48, and 72 hrs of exposure. The average specific growth rate, and the area under the growth curves was then calculated for the exposure period. Significant reduction in growth as compared to controls was seen in the 3 and 10 mg/l test concentrations. The 72 -hr ErL50 for R. subcapitata was calculated to be 2.9 mg/l (WAF), and the 72 -hr EbL50 is 2.6 mg/l (WAF).

Description of key information

There is data available for this substance.Key data is summarized below.

The data used to characterize the acute aquatic toxicity of hydrocarbons, C9, aromatics are from toxicity study with Pseudokirchneriella subcapitata, following standard test guidelines.

P. Subcapitata was exposed to 0, 0.1, 0.4, 1, 3, or 10 mg/l (WAF) of the test substance for 72 hrs. The results indicate that the 72 -hr ErL50 for P. subcapitata was calculated to be 2.9 mg/l (WAF), and the 72 -hr EbL50 is 2.6 mg/l (WAF).

Key value for chemical safety assessment

Additional information

The data used to characterize the acute aquatic toxicity of hydrocarbons, C9, aromatics are from mulitple short-term toxicity studies withPseudokirchneriella subcapitata, following standard test guidelines.

The results indicate that hydrocarbons, C9 aromatics cause moderate acute toxicity to freshwater green algae at a range of 2.9 to ≥ 7.9 mg/L (growth rate), and 2.6 to ≥ 3.8 mg/L (biomass), based on nominal loading of the test substance in water

 

In the WAF methodology, different loadings of the substance are added to the test medium and equilibrium between the water and the hydrocarbons is achieved according to their specific water solubility. The hydrocarbon concentrations at equilibrium will be characteristic of the loading of the substance, and therefore toxicity values from WAF studies are expressed as effect loadings or lethal loadings (EL or LL), not as concentrations. These effect loadings are used to determine environmental classifications. The WAF methodology is widely accepted for the testing of complex hydrocarbon substances and other UVCBs, and it has been incorporated in different guidance documents, including the REACH guidance.