4-(1,4-dioxa-spiro[4.5]dec-8-yl)-cyclohexanone

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1997-11-24 till 1998-02-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

Adopted 17 July, 1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

EEC directive 92/69

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples were taken form 0, 18, 56 and 180 mg/L for analysis at start and end of test. 10 mL were taken from the approximate center of the test vessels. All samples were stored in a deep-freez unit until analysis.

Vehicle:

no

Details on test solutions:

The test substance was soluble at the concentrations tested. Stock solutions of 100 mg/L were used to prepare the lower concentrations. A solution containing 180 mg/L was separately prepared. Solutions were stirred for ten to twenty minutes, in addition with ultra-sonic treatment applied.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Strain: Teleostei, Cyprinidae (Linnaeus, 1758)
- Source: Zodiac, preofacc, De Haar Vissen, L.U. Wageningen, The Netherlands
- Length at study initiation: 2.18 ± 0.13 cm
- Weight at study initiation: 0.38 ± 0.04 g

ACCLIMATION
- Acclimation period: at least 12 days
- Acclimation conditions: same as test conditions
- Type of food during acclimation: Trouvit
- Feeding frequency during acclimation: Daily
- Health during acclimation: mortality was less than 5 %.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

250 mg CaCO3/L

Test temperature:

20.2 - 20.7 °C

pH:

7.6 - 8.8

Dissolved oxygen:

6.2 - 9.4

Nominal and measured concentrations:

Nominal concentrations: 18, 32, 56, 100 and 180 mg/L
Mean measured concentrations: 17.15, not measured, 53.6, not measured and 182 mg/L. Details in Table 3 below.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass vessels
- Material, fill volume: glass, 4L
- Aeration: not in the first 24 h of the test, thereafter test solutions were gently aerated until end of test
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Biomass loading rate: 0.67 g fish/L, i.e. 7 fish per 4 L of test medium

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: ISO medium according to OECD 203

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h photoperiod daily
- Light intensity: not available

EFFECT PARAMETERS MEASURED
at 2.5, 24, 48, 72 and 96 h mortality and other effects on fish were measured. Dead fish were removed when oberved.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0.1, 1, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

pentachlorophenol (PCP)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 180 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

56 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: swimming capacity

Details on results:

- Behavioural abnormalities and mortalities: yes, see Table 1 and Table 2
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

Validation criteria:
- No mortality was observed in the control.
- The analytical program provided clear evidence that the actual concentration had been maintained at more than 80 % of the initial concentrations.
- Further, all test conditions remained within the ranges prescribed by the protocol.

Results with reference substance (positive control):

The LC50 of the reference substance was 0.14 mg/L (95% fiducial limits: 0.13-0.18 mg/L) after 96h. The range of the 96h-LC50 for carp is generally between 0.10 and 0.46 mg/L based on historical data of reference test approximately every 3 month since April 1988. The carp originating from this batch can be considered as sensitive to toxic substances.

Sublethal observations / clinical signs:

Table 1: Specification of effects observed during the study

Concentration (mg/L)	Time (hour)	Specification of effects	Relative number
0-56	0-96	No abnormalities	7/7
100	2.5	Hypoactive	7/7
	24-48	Hypoactive and discolored	7/7
	72	Hypoactive	5/7
		Hypoactive and discolored	2/7
	96	Hypoactive	6/7
		Hypoactive and discolored	1/7
180	2.5	Hypoactive	5/7
		Immobile	2/7
	24	Hypoactive and discolored	5/5
	48	Hypoactive and discolored	1/5
		Immobile and discolored	4/5
	72-96	Hypoactive and discolored	4/5
		Immobile and discolored	1/5

Table 2: Incidence and total mortality

Concentration (mg/L)	Initial number of fish	Cumulative mortality					Total Mortality (%)
		2.5h	24 h	48 h	72 h	96 h
0 (control)	7	0	0	0	0	0	0
18	7	0	0	0	0	0	0
32	7	0	0	0	0	0	0
56	7	0	0	0	0	0	0
100	7	0	0	0	0	0	0
180	7	0	2	2	2	2	29

 

Table 3: Concentrations of test item in test medium

Time [hours]	Concentrations [mg/L]
	Nominal	Analysed*	Relative to nominal [%]
0	0	n.d.	n.a.
	18	18.7	104
	56	59	105
	180	191	106
96	0	n.d.	n.a.
	18	15.6	86
	56	48.3	86
	180	173	96

*Mean of duplicate analysis. The maximum deviation between the responses was calculated tor each sample and was < 10%.

n.d. not detected

n.a. not applicable

Validity criteria fulfilled:

yes

Conclusions:

The test substance has no acute toxicity effect on carp under the present conditions of the test up 100 mg/L, LC50>100 mg/L.

Executive summary:

The acute toxicity of the test item in Carp was performed according to EU Method, Part C.1. and the OECD guideline No. 203. After a range-finding test, a final test was executed with cyprinus carpio exposed to the test item in concentrations of 18, 32, 56, 100 and 180 mg/L in a static system. Seven carp were exposed per concentration and a control. Samples for analysis were taken at the start and the end of the test. Analysis of the samples taken showed that the measured concentrations were in agreement with the nominal concentrations (less than 20% deviation). In the control and the lower concentrations no mortality was observed, only at 180 mg/L two fish died after 24 h (mortality of 29%). Effects on swimming capacity and hypoactivity were recorded at 100 and 180 mg/L. The test item induced no lethal effects in carp at or below 100 mg/L and no sublethal effects at or below 56 mg/L. The 96h-LC50 is > 100 mg/L.

Description of key information

The test substance has no acute toxicity effect on carp under the present conditions of the test up 100 mg/L, LC50>100 mg/L (reference 6.1.1 -1).

Additional information

The acute toxicity of the test item in Carp was performed according to EU Method, Part C.1. and the OECD guideline No. 203. After a range-finding test, a final test was executed with cyprinus carpio exposed to the test item in concentrations of 18, 32, 56, 100 and 180 mg/L in a static system. Seven carp were exposed per concentration and a control. Samples for analysis were taken at the start and the end of the test. Analysis of the samples taken showed that the measured concentrations were in agreement with the nominal concentrations (less than 20% deviation). In the control and the lower concentrations no mortality was observed, only at 180 mg/L two fish died after 24 h (mortality of 29%). Effects on swimming capacity and hypoactivity were recorded at 100 and 180 mg/L. The test item induced no lethal effects in carp at or below 100 mg/L and no sublethal effects at or below 56 mg/L. The 96h-LC50 is > 100 mg/L (reference 6.1.1 -1).