(3E)-dec-3-en-2-one

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:

June 27, 2011 to September 21, 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Freshly prepared samples for analysis were collected at the start of the test and at the test medium renewal at day 1, day 2 and day 3; the aged test media samples were collected at day 1, day 2, day 3 and at day 4.
The freshly prepared samples from the start of the test and from the test medium renewal at day 3, as well as the aged test media samples from day 1 and from day 4 were analysed by gas chromatography.
All samples were analysed directly after sampling without prior storage.
Approximately 50 mg test item were diluted with 50 mL acetonitrile to obtain a stock solution of 1 g test item/L. Appropriate amounts of the stock solution were diluted with culture medium to obtain fortified samples at concentrations of 0.1, 0.2, 1.0 and 3 mg test item/L.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution of nominally 100 mg test item/L was prepared by dissolving 113.6, 109, 93.8 and 94.5 mg test item into 1136, 1090, 938, and 945 mL test water, respectively, by intense stirring for 20 minutes. Then, adequate volumes of this stock solution were mixed into test water to obtain the desired test concentrations.
The test media were prepared just before introduction of the test fish (= start of the test and test medium renewal on day 1, 2 and 3).
- Eluate: n.a.
- Differential loading: n.a.
- Controls: medium without test item
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): n.a.
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): n.a.
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): There were no remarkable observations on the appearance of the test item in test medium
- Other relevant information: A semi static test procedure was chosen with a test medium renewal every 24 h to keep the concentrations of (3E)-dec-3-en-2-one in the test medium as constant as possible during the test period. During this test surviving fish were placed in a clean aquarium with freshly prepared test medium of the corresponding concentration every day. The test media were slightly aerated during the test period.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Forellenzuchtbetrieb Stork, 88348 Bad Saulgau, Germany.
- Age at study initiation (mean and range, SD): Juveniles
- Length at study initiation (length definition, mean, range and SD): 4.80 cm ± 0.3 cm (Mean ± SD)
- Weight at study initiation (mean and range, SD): 1.08 ± 0.29 g (Mean ± SD)
- Method of breeding: All fish were obtained and held in the laboratory for at least 12 days before the start of the test. They were held in water of the quality to be used in the test for at least seven days immediately before testing under the following conditions:
- Light: 12 - 16 hours photoperiod daily
- Temperature: 13 - 17 °C
- Oxygen concentration: at least 80 % of the air saturation value
- Feeding: three times per week or daily until 24 hours before the test was started.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

14 - 15 °C

pH:

7.9 - 8.1

Dissolved oxygen:

91 - 99% of the air saturation value

Nominal and measured concentrations:

nominal: 3.0, 1.36, 0.62, 0.28 and 0.13 mg test item/L and a control.
The 3 highest treatment groups correspond to the following geometric mean measured concentrations of the test item: 2.22, 1.02 and 0.45 mg test item/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria
- Material, size, headspace, fill volume: 12 L
- Aeration: Yes
- Renewal rate of test solution (frequency/flow rate): every 24 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Analytical grade salts were added to deionised water (conductivity <= 10 µS.cm-1) to give the following nominal concentrations:
- CaCl2 x 2H2O : 2.0 mmol/L (= 294.0 mg/L)
- MgSO4 x 7H2O : 0.5 mmol/L (= 123.0 mg/L)
- NaHCO3 : 0.75 mmol/L (= 65.0 mg/L)
- KCl : 0.075 mmol/L (= 5.8 mg/L)
- Water Hardness : 2.5 mmol/L (= 250.0 mg/L) as CaCO3
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4 : 1
- Culture medium different from test medium: No
- Intervals of water quality measurement: The water temperature, pH-values and the dissolved oxygen concentrations were determined daily in the freshly prepared and aged test media of each treatment group.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 h light : 8 h dark
- Light intensity: 560 to 890 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The test fish were observed after approximately 2, 24, 48, 72 and 96 hours test duration for sublethal effects and mortality. Dead fish were removed at least once daily and discarded.
The appearance of the test item in the test medium was observed at least once every day in all freshly prepared and aged test media of all test concentrations test concentration.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2
- Range finding study
- Results used to determine the conditions for the definitive study: Yes

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.5 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.:

1.02 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

In the control and at concentrations up to and including nominal 0.62 mg test item/L all fish survived until the end of the experiment and there were no signs of intoxication. At the highest test item concentration of nominally 3.0 mg/l (corresponding to a geometric mean measured concentration of 2.22 mg test item/L) all fish showed sublethal effects in terms of strong ventilation, dark colouration, extended gills, tumbling swimming and they were lying on their side or back after 2 hours of exposure. After 24 hours of exposure 4 of the 7 animals were dead and the others were lying on their side or back, tumbled during swimming and had extended gills and strong ventilation. After 48 hours of exposure all fish at nominally 3.0 mg test item/L were dead. At the second highest concentration 0f nominally 1.36 mg test item/L (corresponding t0 a geometric mean measured concentration of 1.02 mg test item/L) all fish displayed sublethal effects from 24 hours of exposure onwards until the end of the test. These effects included strong ventilation, tumbling swimming, extended gills, dark colouration and exophthalmos. Additionally the fish were mainly located in the bottom of the aquarium lying on their side or back.

Reported statistics and error estimates:

The LC50 at the observation times was calculated by Probit analysis using the geometric mean measured test item concentrations of the 3 highest treatment groups since the lower concentrations were below LOQ and it was not possible to calculate correct mean measured test concentrations. No mortality or sublethal effects occurred in the two lower concentration groups therefore they were not necessary for the statistical evaluations. The NOEC, the LOEC, the LC0 and the LC100 were determined directly from the raw data.

Sublethal observations / clinical signs:

Table 1 . Observed Mortality of unfed Rainbow Trout (Oncorhynchus mykiss) exposed to (3E)-dec-3-en-2-one for 96 hours in a Static Test

Mortality
Nominal Concentration	Measured concentration	Exposure Time [h]
[mg/L]	[mg/L]	0	2	24	48	72	96
		# mort	# mort	# mort	# mort	# mort	# mort
Control	Control	0	0	0	0	0	0
0.13	n.d.*	0	0	0	0	0	0
0.28	n.d.*	0	0	0	0	0	0
0.62	0.45	0	0	0	0	0	0
1.36	1.02	0	0	0	0	0	0
3.0	2.22	0	0	4	7	7	7
LC50 [mg/L]		n.d.	n.d.	2.16	1.5	1.5	1.5
95% C.I.		n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
# mort: Number of dead fish
n.d.: could not be determined
n.d.*: values could not be calculated since the analytical results are below LOQ or LOD
C.I.: Confidence intervals LC50 values refer to geometric mean measured test concentrations.

Table 2. Mortality and Sublethal Effects

Mortality and Sublethal Effects
Nominal concentration [mg test item/L]
Exposure Time [h]	Control	0.13	0.28	0.62	1.36	3
#mort 0 h    #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 0 -	0 0 -
#mort 2 h    #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, SB, SV, TS
#mort 24 h  #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 EG, MB, SV	0 7 EG, SB, SV, TS
#mort 48 h  #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, SB, SV, TS	7 - -
#mort 72 h #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, SB, SV	7 - -
#mort 96 h  #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, EX, SB, SV, TS	7 - -
# mort: Number of dead fish
# symp: Number of fish With sublethal effects
DC: dark colouration
EG: strongly extended gills
EX: exophthalmus
MB: fish mainly on the bottom
SB: fish lying on side or back on the bottom
SV: hyperventilation
TS: tumbling during swimming

Table 3. Dissolved Oxygen Concentrations (%) in the Test Media

Mortality and Sublethal Effects
Nominal concentration [mg test item/L]
Exposure Time [h]	Control	0.13	0.28	0.62	1.36	3
#mort 0 h    #symp          symp	0 0 -	0 0 -	0 0 -	0 0 -	0 0 -	0 0 -
#mort 2 h    #symp          symp	0 0 -	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, SB, SV, TS
#mort 24 h  #symp          symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 EG, MB, SV	0 7 EG, SB, SV, TS
#mort 48 h  #symp          symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, SB, SV, TS	7 - -
#mort 72 h #symp         symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, SB, SV	7 - -
#mort 96 h  #symp          symp	0 0 -	0 0 -	0 0 -	0 0 -	0 7 DC, EG, EX, SB, SV, TS	7 - -
# mort: Number of dead fish
# symp: Number of fish With sublethal effects
DC: dark colouration
EG: strongly extended gills
EX: exophthalmus
MB: fish mainly on the bottom
SB: fish lying on side or back on the bottom
SV: hyperventilation
TS: tumbling during swimming

Table 4. Appearance of the Test Item in Test Medium

Nominal Concentration	Eposure Time [h]
[mg/l]		0 h	2 h	24 h	48 h	72 h	96 h
Control	new age	0 -	- 0	0 0	0 0	0 0	- 0
0.13	new age	0 -	- 0	0 0	0 0	0 0	- 0
0.28	new age	0 -	- 0	0 0	0 0	0 0	- 0
0.62	new age	0 -	- 0	0 0	0 0	0 0	- 0
1.36	new age	0 -	- 0	0 0	0 0	0 0	- 0
3.0	new age	0 -	- 0	0 0	- 0	- -	- -

Abbreviations:

0: No remarkable observations, clear test medium

1: Colouration caused by the test item

2: Turbidity caused by the test item

3: Inhomogeneous dispersion of the test item

4: Precipitation of the test item

5: Test item at the surface

6: Test item lying at the bottom of the test vessel

7: Test item caused foaming

Validity of the test:

- In the control the mortality did not exceed 10 % at the end ofthe test.

- The dissolved oxygen concentration in the test media did not fall below 60 % of air saturation value during the test.

Validity criteria fulfilled:

yes

Conclusions:

In a 96-h acute toxicity study, Rainbow Trout (Oncorhynchus mykiss) were exposed to 3 -decen-2 -one at nominal concentrations of 0.0 (control), 0.13, 0.28, 0.62, 1.36 and 3.0 mg/L (the three highest treatment groups correspond to the following geometric mean measured concentrations of the test item: 0.45, 1.02 and 2.22 mg test item/L) under semi-static conditions (renewal every 24 h). 
The following effect levels were determined:
96-hour LC50: 1.50 mg test item/L (geometric mean measured); 95 % Confidence Interval: n.d.;
96-hour LC0: 1.02 mg test item/L (geometric mean measured).

Executive summary:

In a 96-h acute toxicity study, rainbow trout (Oncorhynchus mykiss) were exposed to 3-decen-2-one at nominal concentrations of 0 (control), 0.13, 0.28, 0.62, 1.36, and 3.0 mg/L under semi-static conditions (renewal every 24 h). The three highest treatment groups correspond to the following geometric mean measured concentrations of the test item: 0.45, 1.02 and 2.22 mg test item/L. The nominal concentrations of 0.13 and 0.28 could not be verified analytically as the measurements were below the LOQ of the method (0.2 mg test item/L). The 96-h LC50 was 1.5 mg/L. The LC0 value, based on mortality, were 1.02 mg/L, respectively.

Since the deviation from the nominal concentrations are greater than 20 per cent, all results are based on the geometric mean measured concentrations.

No mortality or sublethal effects occurred in the two lower concentration groups. In the control and at concentrations up to and including nominal 0.62 mg test item/L all fish survived until the end of the experiment and there were no signs of intoxication. At the highest test item concentration of nominally 3.0 mg/L (corresponding to a geometric mean measured concentration of 2.22 mg test item/L) all fish showed sublethal effects in terms of strong ventilation, dark colouration, extended gills, tumbling swimming and they were lying on their side or back after 2 hours of exposure. After 24 hours of exposure 4 of the 7 animals were dead and the others were lying on their side or back, tumbled during swimming and had extended gills and strong ventilation. After 48 hours of exposure all fish at nominally 3.0 mg test item/L were dead. At the second highest concentration of nominally 1.36 mg test item/L (corresponding to a geometric mean measured concentration of 1.02 mg test item/L) all fish displayed sublethal effects from 24 hours of exposure onwards until the end of the test. These effects included strong ventilation, tumbling swimming, extended gills, dark colouration and exophthalmos. Additionally, the fish were mainly located in the bottom of the aquarium lying on their side or back.

Based on the results of this study, 3-decene-2-one would be classified regarding the aquatic hazard as chronic category 2 to O. mykiss in accordance with the Regulation (EC) No 1272/2008. The substance has not to be categorized as acute toxic to the aquatic environment.

This toxicity study is considered as acceptable and satisfies the guideline requirement for an acute fish toxicity study.

Results synopsis

Rainbow trout (Oncorhynchus mykiss), Juveniles, length 4.80 ± 0.3 cm

Test type: semi-static

96-hour LC50: 1.50 mg test item/L (geometric mean measured); 95 % Confidence Interval: n.d.;

96-hour LC0: 1.02 mg test item/L (geometric mean measured).

Endpoint effected: Survival/mortality

Description of key information

In a 96-h acute toxicity study, rainbow trout (Oncorhynchus mykiss) were exposed to (3E)-dec-3-en-2-one at nominal concentrations of 0 (control), 0.13, 0.28, 0.62, 1.36, and 3.0 mg/L under semi-static conditions (renewal every 24 h). The three highest treatment groups correspond to the following geometric mean measured concentrations of the test item: 0.45, 1.02 and 2.22 mg test item/L.

The following effect levels were determined:

96-hour LC50: 1.50 mg test item/L (geometric mean measured); 95 % Confidence Interval: n.d.;

96-hour LC0: 1.02 mg test item/L (geometric mean measured).

No mortality or sublethal effects occurred in the two lower concentration groups. In the control and at concentrations up to and including nominal 0.62 mg test item/L all fish survived until the end of the experiment and there were no signs of intoxication.

At the nominal concentration 1.36 mg test item/L all fish displayed sublethal effects from 24 hours of exposure onwards until the end of the test. These effects included strong ventilation, tumbling swimming, extended gills, dark colouration and exophthalmos. Additionally, the fish were mainly located in the bottom of the aquarium lying on their side or back. At the highest nominal test item concentration of 3.0 mg/L all fish showed sublethal effects in terms of strong ventilation, dark colouration, extended gills, tumbling swimming and they were lying on their side or back after 2 hours of exposure. After 24 hours of exposure 4 of the 7 animals were dead and the others were lying on their side or back, tumbled during swimming and had extended gills and strong ventilation. After 48 hours of exposure all fish were dead.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.5 mg/L