Isopropyl acetoacetate

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:

1989-03-13 to 1989-04-14

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: April 04, 1984

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

April 25, 1984

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 0*, 22, 25, 50, 100, 220 and 500 mg/L (0* Solvent Control with 0.095 mL Tween 80/L)

- Sampling method: samples were determined at the beginning of the test, after 48 and 96 hours.

Vehicle:

yes

Remarks:

Tween 80

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The test substance was mixed with 0.095 mL Tween 80/L as a solubiliser, diluted with dilution water, homogenised in an ultrasonic bath and introduced into the test tanks while stirring with a glass rod. At 500 mg/L slight substance deposits on the water surface and at 100 mg/L substance deposits on the bottom of the basin could be observed.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Strain: Brachydanio rerio (HAMILTON-BUCHANAN)
- Source: West Aquarium, 3422 Bad Lauterberg, Germany
- Length at study initiation: 2.5 – 3.5 (mean = 2.9 and SD = 0.28)

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: Tetra Min, ad libitum
- Feeding frequency during acclimation: daily ad libitum

FEEDING DURING TEST: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Concentration group: 21.0 °C – 22.8 °C;
Control: 21.0 °C – 21.9 °C

pH:

Concentration group: 7.5 – 8.2;
Control: 7.6 – 8.2

Dissolved oxygen:

Concentration group: 6.7 mg/L – 9.4 mg/L;
Control: 7.0 mg/L – 9.3 mg/L

Nominal and measured concentrations:

Nominal Concentrations:0, 0*, 22, 50, 100, 220 and 500 mg/L (0* Solvent Control with 0.095 mL Tween 80/L);

Measured concentrations: The analytical values averaged over 80 % of the nominal concentration.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass
- Type: open
- Material, fill volume: glass, 100 L
- Aeration: yes
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1

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

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 12 hours light – 12 hours dark
- Light intensity: 700 Lux

EFFECT PARAMETERS MEASURED: The fish were observed for signs of toxicity or death after 24, 48, 72 and 96 hours.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

220 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

220 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 50 - <= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

>= 50 - <= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

A probit analysis was not possible due to the present lethality rate. Since the factor of the concentration levels was 2.2 (3rd root of10), no further concentrations were tested. The LC50 was given as a range.

Sublethal observations / clinical signs:

To determine the substance content, at the beginning of the experiment, after 48 and 96 hours, approx. 100 ml were taken from the centre of the basin with a glass screw bottle

The averaged values were as follows:

Nominal (mg/L)	Mean (mg/L)	%
50	49.3	98.5
100	98.5	98.5

The analytical values were averaged over 80 % of the nominal concentration. Only two concentrations were measured due to the water solubility of the test item.

Content of substances in water

Nominal Concentration (mg/L)	Amount recovered after the start of test (mg/L)
	0 h	48 h	96 h
50	49.8
50		49.2
50			48.8
100	100
100		97.9
100			97.7

Validity criteria fulfilled:

yes

Conclusions:

The toxicity of the test item to Danio rerio was determined by a static test. Under the given conditions of this study, the 96 h LC50 value was in the range of 50 – 100 mg/L and the 96 h LC0 value was determined to be 50 mg/L.

Executive summary:

The objective of this study was to determine the acute fish toxicity of the test material using zebrafish (Danio rerio) according to OECD TG 203 (1984) under GLP conditions. For this purpose, 10 zebrafish per test item group and 10 zebrafish in a control group were exposed over 96 hours under defined conditions. The fish were observed for symptoms of toxicity or death. Zebrafish were exposed to aqueous test item nominal concentration of 22, 50, 100, 220 and 500 mg/L in an open static test system. Tween 80 was used as a solubilizer due to the test item's dissolution behaviour in the test medium. Additionally, a solvent control with 0.095 mL Tween 80/L was tested. At 100 and 500 mg/L substance deposits could be observed. Samples were taken from the 50 and 100 mg/L concentration groups and were analysed using HPLC with UV-detection. To determine the substance content, water samples were taken from the middle of the basin at the beginning of the test, after 48 and 96 hours. The analytical values were averaged over 80 % of the target concentration. Therefore, the biological endpoints were based on the nominal concentrations and the LC₅₀ was given as a range. No signs of toxicity were seen in the control group. The zebrafish exposed to concentrations of 22 and 50 mg/L showed symptoms but no lethality. Symptoms of toxicity and lethality were seen in the concentrations 100, 220, and 500 mg/L. Under the given conditions of this study, the 96 h LC₅₀ value was in the ranged 50 – 100 mg/L. the LC₀ was determined to be 50 mg/L.

Description of key information

The toxicity of the test item to Danio rerio was determined by a static test. Under the given conditions of this study, the 96 h LC₅₀ value was a range of 50 – 100 mg/L and the 96 h LC₀ value was determined to be 50 mg/L (reference 6.1.1-1).

Additional information

The objective of this study was to determine the acute fish toxicity of the test material using zebrafish (Danio rerio) according to OECD TG 203 (1984) under GLP conditions. For this purpose, 10 zebrafish per test item group and 10 zebrafish in a control group were exposed over 96 hours under defined conditions. The fish were observed for symptoms of toxicity or death. Zebrafish were exposed to aqueous test item nominal concentration of 22, 50, 100, 220 and 500 mg/L in an open static test system. Tween 80 was used as a solubilizer due to the test item's dissolution behaviour in the test medium. Additionally, a solvent control with 0.095 mL Tween 80/L was tested. At 100 and 500 mg/L substance deposits could be observed. Samples were taken from the 50 and 100 mg/L concentration groups and were analysed using HPLC with UV-detection. To determine the substance content, water samples were taken from the middle of the basin at the beginning of the test, after 48 and 96 hours. The analytical values were averaged over 80 % of the target concentration. Therefore, the biological endpoints were based on the nominal concentrations and the LC₅₀ was given as a range. No signs of toxicity were seen in the control group. The zebrafish exposed to concentrations of 22 and 50 mg/L showed symptoms but no lethality. Symptoms of toxicity and lethality were seen in the concentrations 100, 220, and 500 mg/L. Under the given conditions of this study, the 96 h LC₅₀ value was in the ranged 50 – 100 mg/L. the LC₀ was determined to be 50 mg/L.