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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Guideline study, comparable to OECD 203. Scientifically acceptable.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1988
Report date:
1988

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
other: German standard DIN 38 412 L15
GLP compliance:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
2,5,6-trimethylcyclohex-2-en-1-one
EC Number:
243-473-0
EC Name:
2,5,6-trimethylcyclohex-2-en-1-one
Cas Number:
20030-30-2
Molecular formula:
C9H14O
IUPAC Name:
2,5,6-trimethylcyclohex-2-en-1-one
Details on test material:
- Name of test material (as cited in study report): 2,3,6-Trimethylcyclohexen-1-one
- Analytical purity: about 95 %
water solubity: < 1000 mg/l

Sampling and analysis

Analytical monitoring:
no

Test solutions

Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test substance was added to the test water without any pretreatment. Then, the fish were placed into the test aquaria.

Test organisms

Test organisms (species):
Leuciscus idus
Details on test organisms:
TEST ORGANISM
- Common name: Golden orfe
- Source: Fischzucht Paul Eggers, Hohenweststedt, Germany
- Length at study initiation (length definition, mean, range and SD): 7.7 cm (range 6.9 - 8.9 cm)
- Weight at study initiation (mean and range, SD): 4.3 g (range 2.8 - 6.6 g)
- Corpulence factor of the test fish batch: 0.9
- Housing and adaption: Culture conditions: oil-free aerated and charcoal filtered tap water, flow-through system; Water temperature: 13 - 22 °C Duration of housing: about 4 month Mortality during the time of housing: 0 %, Medical treatment: 4 times with 0.05 mg/l malachite green chloride, twice with 10 mg/l tetracycline hydrochloride
- Feeding during test: 1 day before and during exposure.


ACCLIMATION
- Acclimation period: 3 days
- Type and amount of food: Growing feed F/B 50, ad libitum.
- Health during acclimation (any mortality observed): no mortality observed

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h

Test conditions

Hardness:
2.5 mmol/l
Test temperature:
20 - 21 °C
pH:
7.6 - 8.0
Dissolved oxygen:
7.6 - 8.7 mg/l
Nominal and measured concentrations:
Nominal test concentrations: 4.64, 10.0, 21.50, 46.40, 100.0 mg/l (based on the results of a range-finding test). In addition, a control was tested in parallel.
Details on test conditions:
TEST SYSTEM
- Test vessel: All glass aquaria (30 x 22 x 24 cm)
- Type (delete if not applicable): open / closed
- Fill volume: 10 l
- Aeration: slight
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 4.3 g fish/l test water


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted freshwater according to DIN 38412, Part II, 1982; prepared from fully demineralised tap water (conductivity: max 10 micro MHO) by adding: 294.0 mg/l CaCl2 * 2 H2O, 123.3 mg/l MgSO4 + 7 H2O, 63.0 mg/l NaHCO3, 5.5 mg/l KCl
- Ratio Na/K ions: 10 : 1
- Acid capacity: 0.8 mmol/l
- Ca/mg ratio: 4 : 1

OTHER TEST CONDITIONS
- Photoperiod: 16 : 8 hours day-night regime

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and symptoms of intoxication after 1, 4, 24, 48, 72 and 96 h.

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
21.5 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
46.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
68.12 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Results with reference substance (positive control):
48 h-LC50 of the positive control Chloroacetamide: about 26 mg/l (corresponds to normal sensitivity).
Reported statistics and error estimates:
The NOEC, the LOEC, the LC0 and the LC100 were determined directly from the raw data. The LC50 was determined from the geometrical mean of the LC0 (46.4 mg/l) and the LC100 (100 mg/l).

Any other information on results incl. tables

Sublethal observations / clinical signs:
During the 96 h exposure to 2,3,6-Trimethylcyclohexen-1-on mortalities were as follows:

Nominal test Cumulative mortalitiy (%) after x hours
concentration 1 / 24 / 48 / 72 / 96
(mg/l)
Control 0 / 0 / 0 / 0 / 0
4.64 0 / 0 / 0 / 0 / 0
10.0 0 / 0 / 0 / 0 / 0
21.5 0 / 0 / 0 / 0 / 0
46.4 0 / 0 / 0 / 0 / 0
100.0 8 / 10 / 10 / 10 / 10

Symptoms of intoxication:

Nominal test Symptoms of intoxication after x hours
concentration 1 / 24 / 48 / 72 / 96
(mg/l)
Control - / - / - / - / -
4.64 - / - / - / - / -
10.0 - / - / - / - / -
21.5 - / - / - / - / -
46.4 A / A / A / A / +
100.0 N / + / + / + / +


Definitions of Symptoms:
-: No symptoms
+: All fish dead
N: Narcotic-like state
A: Apathy

Behavior of the test item during the test:
No remarkable observations.

Applicant's summary and conclusion

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