2-hydroxy-3-(prop-2-enoyloxy)propyl 2-methyl-2-propylhexanoate

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:

From October 16, 2016 to November 11, 2016

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:

yes

Remarks:

on the study plan but uncritical

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

yes

Remarks:

on the study plan but uncritical

Principles of method if other than guideline:

Corresponding SOP of LAUS GmbH: SOP 118 00 504 edition 4, valid from 01. October 2013 „Bestimmung der akuten Wirkung von Substanzen auf Zebrabärblinge (Danio rerio)“.

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch no.: JBGJ0045R
Purity: 100% (UVCB)
Appearance: clear yellowish liquid

Analytical monitoring:

yes

Remarks:

HPLC-UV

Details on sampling:

In the first experiment the content of the test substance was determined using HPLC-UV determination at the start of test. As it was aborted after 48 hours and no further analytically determinations were performed.
In the second experiment the content of the test substance in the freshly prepared and aged test solutions was determined using HPLC-UV -determination at the start and at the end of the test and at medium renewal after 48 hours. Because of the limited solubility of the test substance, the correlation between nominal and measured concentration was weak. Therefore, the determination of the biological results was based on the geometric mean of the measured concentrations.

Vehicle:

no

Details on test solutions:

The water-accommodated fractions (WAF) were prepared for the test.
In the first experiment this was done by pipetting 46, 22, 10, 4.6 and 2.2 µL/L corresponding to the nominal loads 46, 22, 10, 4.6 and 2.2 mg/L (based on a density of 1.01 g/mL, stated in the MSDS provided by the sponsor) on the surface of dilution water and stirring slowly for 24 hours. The lower phase was used as test solution.
In the second experiment the water-accommodated fractions (WAF) were prepared by pipetting 3.2, 1.8, 1.0, 0.56 and 0.32 µL/L corresponding to the nominal loads 3.2, 1.8, 1.0, 0.56 and 0.32 mg/L on the surface of dilution water and stirring slowly for 24 hours. The lower phase was used as test solution.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Danio rerio HAMILTON BUCHANAN from Umweltbundesamt (UBA) (1rst experiment) and Dehner Garten Center, 67433 Neustadt (2nd experiment)
Length: 2 +/- 1 cm
Vessels: polyethylene aquaria
Medium: chlorine-free tap water
Feeding: three times a day with warmwater fishfood
Photo period: 12/12 hours, using neon tubes

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

-

Test temperature:

23 +/- 2°C

pH:

7.9 - 9.1

Dissolved oxygen:

7.9 - 9.1 mg/L O2

Salinity:

-

Conductivity:

-

Nominal and measured concentrations:

Nominal concentrations:
First experiment: 46, 22, 10, 4.6, 2.2 and 0.0 mg/L
Second experiment: 3.2, 1.8, 1.0, 0.56, 0.32 and 0.0 mg/L

Details on test conditions:

First experiment:
Duration: 48 hours
Loading: 1 fish/L
Vessels: glass aquaria, maximal volume 10 L
Aeration: accomplished with glass tubes
Feeding: none
Photo period: 12/12 hours using neon tubes
Temperature: 22.3 – 22.5°C
pH adjustment: none
Replicates (treatments): 1 vessel, each containing 7 L test solution and 7 fish
Replicates (blank Control): 1 vessel, containing 7 L tap water and 7 fish
- Observations were made every 24 hours until abortion of the experiment. The values of pH and O2-concentration were documented in each vessel at 0 hours and 24 hours. Mortalities or abnormal behaviour were documented in each vessel every 24 hours until abortion of the experiment. The content of the test substance in the test vessels was measured at the beginning of the test.

Second experiment:
Duration: 46 hours
Loading: 1 fish/L
Vessels: glass aquaria, maximal volume 10 L
Aeration: accomplished with glass tubes
Feeding: none
Photo period: 12/12 hours using neon tubes
Temperature: 22.3 – 22.5°C
pH adjustment: none
Replicates (treatments): 1 vessel, each containing 7 L test solution and 7 fish
Replicates (blank Control): 1 vessel, containing 7 L tap water and 7 fish
Observations were made every 24 hours, measuring pH and O2-concentration of the test solution in each vessel and documenting mortalities or abnormal behaviour. The content of the test substance in the test vessels was measured at the beginning and at the end of the test and at medium renewal after 48 hours in the new and the old solutions.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: measured concentration: 0.86 mg/L

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: measured concentration: 2.02 mg/L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: measured concentration: 1.3 mg/L

Details on results:

First experiment: after 48 hours, all the fish (with the exception of 1 animal in the lowest concentrated treatment) were found dead. Therefore, the experiment was aborted and a second experiment using lower concentrations was conducted.
Second experiment: the recovery of the measured concentration at the start was lower than during validation. This was caused by the presence of the test organism. Mortality was observed only in the highest concentrated treatment. In the other treatments and in the blank control neither mortality nor abnormal behaviour was observed.

The following results for the test substance could be determined:
96h-NOEC = 0.86 mg/L (measured concentration)
96h-LOEC = 2.02 mg/L (measured concentration)
96h-LC50 = 1.3 mg/L (measured concentration)

Additionally, the results based on the nominal concentrations were determined:
96h-NOEC = 1.8 mg/L (nominal concentration)
96h-LOEC = 3.2 mg/L (nominal concentration)
96h-EC50 = 2.4 mg/L (nominal concentration)

Note: NOEC and LOEC were determined by comparing the respective treatment with the blank control. Statistically insignificant variation is considered as “no observed effect”, although the EC values which were read from the graph toxicity vs. concentration may lie lower.

Results with reference substance (positive control):

-

Reported statistics and error estimates:

Statistics: software ToxRat® Professional, version 3. 2.1

Sublethal observations / clinical signs:

During preparation of the WAF at the loading rate 1 mg/L, the magnetic stir bar stopped stirring at an unknown point in time because of a technical defect. Therefore, the measured concentration in this test solution was much too low. Because only in the highest concentrated treatment mortality was observed, treatment 1 mg/L was not necessary for determination of the results. Therefore, the geometric means of the 3 highest measured concentrations (3.2, 1.8 and 0.56 mg/L nominal) were used for determination of the results.

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions, the 96h LC50, LOEC and NOEC values for Danio rerio were determined to be: 1.3, 2.02 and 0.86 mg/L (measured concentrations), respectively (corresponding to nominal concentrations of 2.4, 3.2 and 1.8 mg/L, respectively).

Executive summary:

A study was conducted to determine the short-term toxicity of the test substance to fish according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. Danio rerio were exposed to water accomodated fractions (WAFs) prepared at nominal concentrations of 0, 2.2, 4.6, 10, 22 and 46 mg/L (first experiment) and 0, 0.32, 0.56, 1, 1.8 and 3.2 mg/L (second experiment) under semi-static conditions. The total exposure period was 96 h and samples for analytical confirmation of actual exposure concentrations were taken at the start, at the end and at 48h (following medium renewal) and analysed by HPLC-UV. Mortality and other effects were recorded at 24, 48, 72 and 96 h following the start of exposure. In the first experiment, all the fish (with the exception of 1 animal in the lowest concentrated treatment) were found dead after 48 h. Therefore, the experiment was aborted and a second experiment using lower concentrations was conducted. In the second experiment, the recovery of the measured concentration at the start was lower than during validation. This was caused by the presence of the test organism. Mortality was observed only in the highest concentrated treatment. In the other treatments and in the blank control neither mortality nor abnormal behaviour was observed. Because of the limited solubility of the test substance, the correlation between nominal and measured concentration was weak. Therefore, the determination of the biological results was based on the geometric mean of the measured concentrations. The NOEC and LOEC were determined by comparing the respective treatment with the blank control. Under the study conditions, the 96 h LC50, LOEC and NOEC values for Danio rerio were determined to be: 1.3, 2.02 and 0.86 mg/L (measured concentrations), respectively (corresponding to nominal concentrations of 2.4, 3.2 and 1.8 mg/L, respectively) (Muckle, 2016).

Description of key information

The 96 h LC50 value for Danio rerio due to the test substance was determined to be 1.3 mg/L (measured).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.3 mg/L

Additional information

A study was conducted to determine the short-term toxicity of the test substance to fish according to OECD Guideline 203 and EU Method C.1, in compliance with GLP. Danio rerio were exposed to water accomodated fractions (WAFs) prepared at nominal concentrations of 0, 2.2, 4.6, 10, 22 and 46 mg/L (first experiment) and 0, 0.32, 0.56, 1, 1.8 and 3.2 mg/L (second experiment) under semi-static conditions. The total exposure period was 96 h and samples for analytical confirmation of actual exposure concentrations were taken at the start, at the end and at 48h (following medium renewal) and analysed by HPLC-UV. Mortality and other effects were recorded at 24, 48, 72 and 96 h following the start of exposure. In the first experiment, all the fish (with the exception of 1 animal in the lowest concentrated treatment) were found dead after 48 h. Therefore, the experiment was aborted and a second experiment using lower concentrations was conducted. In the second experiment, the recovery of the measured concentration at the start was lower than during validation. This was caused by the presence of the test organism. Mortality was observed only in the highest concentrated treatment. In the other treatments and in the blank control neither mortality nor abnormal behaviour was observed. Because of the limited solubility of the test substance, the correlation between nominal and measured concentration was weak. Therefore, the determination of the biological results was based on the geometric mean of the measured concentrations. The NOEC and LOEC were determined by comparing the respective treatment with the blank control. Under the study conditions, the 96 h LC50, LOEC and NOEC values for Danio rerio were determined to be: 1.3, 2.02 and 0.86 mg/L (measured concentrations), respectively (corresponding to nominal concentrations of 2.4, 3.2 and 1.8 mg/L, respectively) (Muckle, 2016).