2,6,6-trimethylcyclohex-2-ene-1,4-dione

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,6,6-trimethylcyclohex-2-ene-1,4-dione

EC Number:

214-406-2

EC Name:

2,6,6-trimethylcyclohex-2-ene-1,4-dione

Cas Number:

1125-21-9

Molecular formula:

C9H12O2

IUPAC Name:

2,6,6-trimethylcyclohex-2-ene-1,4-dione

Details on test material:

- Name of test material (as cited in study report): Oxoisophoron
- Lot/batch No.: LJ 28078/V70
- Analytical purity: 98.87 %;

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: the test concentrations were prepared separately.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Source: Charles River Aquatics, Someren, Netherlands
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): 3.32 cm (range 2.5 - 4.2 cm)
- Weight at study initiation (mean and range, SD): 0.203 g (range 0.10 - 0.39 g)


ACCLIMATION
- Acclimation period: 3 days
- Type and amount of food: Tetra Min Standard fish food, Tetra Werke, Melle, Germany; ad libitum
- Feeding frequency: daily
- 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:

about 2.5 mmol/l

Test temperature:

22 °C

pH:

8.1 - 8.4

Dissolved oxygen:

5.8 - 8.6 mg/l

Nominal and measured concentrations:

Nominal test substance concentrations:
0, 21.5, 46.4, 100, 215 and 464 mg/l

Analytically determined test concentrations:
<1, 18.1, 40.6, 86.2, 192.3, 417.3 mg/l

The recovery of the test substance ranged from 98.6 - 102.3 % at test start to 84.2 - 89.9 % at test end.

Details on test conditions:

TEST SYSTEM
- Test vessel: all glass aquaria (30 x 22 x 24 cm)
- Fill volume: 10 l
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.2 g fish/ l test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: charcoal filtered tap water (dechlorinated) from the city of Frankenthal: Continuous aeration with oil-free air

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 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Results used to determine the conditions for the definitive study: LC50 after 96 h >100 <1000 mg/l

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

46.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

behaviour

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

215 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

315.85 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks:

geom. mean

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

464 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Reported statistics and error estimates:

The EC50 value was calculated as the geometrical mean of 215 mg/l (LC0) and 464 mg/l (LC100) value.
Finney, D.J., Probit Analysis, Cambr. Univ. Press, 3rd ed., 1971;

Any other information on results incl. tables

Sublethal observations / clinical signs:

During the 96 h exposure to Oxoisophoron, mortalities were as follows:

Nominal test Cumulative mortalitiy (%) after x hours
concentration 1 / 4 / 24 / 48 / 72 / 96
(mg/l)
Control 0 / 0 / 0 / 0 / 0 / 0
21.5 0 / 0 / 0 / 0 / 0 / 0
46.4 0 / 0 / 0 / 0 / 0 / 0
100 0 / 0 / 0 / 0 / 0 / 0
215 0 / 0 / 0 / 0 / 0 / 0
464 0 / 0 / 10 / 10 / 10 / 10

Symptoms of intoxication:

Nominal test Symptoms of intoxication after x hours
concentration 1 / 4 /24 / 48 / 72 / 96
(mg/l)
Control - / - / - / - / - / -
21.5 - / - / - / - / - / -
46.4 - / - / - / - / - / -
100 (A) / (A) / (A) / - / - / -
215 A / A,C / A / A / A / T,C,D
464 C,D / K,N,T / + / + / + / +

Definitions of Symptoms:
-: No symptoms
+: All fish dead
A: Apathy
N: Narcotic-like state
K: Convulsions
T: Tumbling
C: Swimming near water surface
D: Swimming near bottom
(): slight to very slight effects