Fast Pyrolysis Bio-oil

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:

14 June 2013 – 11 July 2013

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:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Landesanstalt für Umwelt, Messungen und Naturschutz Baden-Württemberg, Germany

Analytical monitoring:

yes

Details on sampling:

Analytical samples from control, 45.5 and 100 mg/L at t = 0, 24, 48 and 72 h from fresh test solutions and at t = 96 h, from aged test solution were analysed.

Vehicle:

no

Details on test solutions:

For the preparation of the test item solutions, the test item was stirred until a homogeneous consistency was obtained. Subsequently an appropriate amount of the test item was directly weighed onto teflon tape and given to test medium and stirred slightly for 48 hours, protected from light, for each individual loading rate. Afterwards the undissolved test item was allowed to sediment and/or float for a period of 24 hours. The test item solution used in the test vessel was withdrawn by suction from the middle of the preparation vessel (according to OECD SERIES ON TESTING AND ASSESSMENT No. 23). Transfer of undissolved test item was avoided. Each solution was applied to the respective test vessel.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

size between 1 - 2 cm; fish from own breeding, according to IME they had been bred from wild strain zebra fish before, only fish in good health and free from any apparent malformation were used. The last feeding was performed two days prior to the test start.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

none

Hardness:

9°dH corresponding to 160 mg CaCO3/L

Test temperature:

23.5 – 24.7 °C

pH:

7.12 – 7.83

Dissolved oxygen:

> 85 % of air saturation

Salinity:

not measured

Nominal and measured concentrations:

nominal test concentrations: 100, 45.5, 20.7, 9.39, 4.27 mg/L (loading rate of the WAF); only the two highest concentrations which did show effects were analysed resulting in measured concentrations of 67.4 mg/ (100 mg/L nominal) and 28.8 mg/L (45.5 mg/L nominal) which corresponds to 67.4 % rsp. 63.4 % of the nominal loading rate

Details on test conditions:

test tanks: 2 L glass vessels filled with 1.5 L test solution; continuous aeration of the test tanks with a membrane pump using a Pasteur pipette, seven fish per test concentration

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

94.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 69.2 – 129 mg/L

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 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.:

63.7 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Remarks on result:

other: 46.6 – 87.1 mg/L

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 67.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

45.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

28.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

28.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

67.4 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

dissolved

Basis for effect:

mortality (fish)

Key result

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

45.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

In the control and in the test item concentrations up to and including 45.5 mg/L no mortality was observed. At the highest test item concentration of 100 mg/L four fish died until the end of the test at 96 h. No sublethal effects were observed during the test in the control and the test item concentrations up to and including 45.5 mg/L. At 100 mg/L three fish showed reduced activity after 48 h, and were found dead after 72 h. After 96 h four fish were found dead at the highest test item concentration. The average weight of the test organisms was 0.0076 +/- 0.0034 g; the average length was 12 +/- 1 mm.

Results with reference substance (positive control):

not performed

Reported statistics and error estimates:

The NOEC (mortality) was established based on the highest test concentration at which the mortality is not higher than the allowed control mortality.
The LC50-value (96 h) was calculated using Spearman-Kärber estimator (ANELLIS & WERKOWSKY, 1968).

Validity criteria fulfilled:

yes

Conclusions:

Fast Pyrolysis Bio-oil (tested as water accommodated fraction) showed only slight toxicity to Danio rerio (LC50, 96 h = 63.7 mg/L, analyzed concentration).

Executive summary:

Short-term toxicity of Fast Pyrolysis Bio-oil (tested as water accommodated fraction) to zebrafish (Danio rerio) was determined in GLP-compliant study according to OECD guideline (No. 203). Mortality was observed at the two highest loading rates (45.5 and 100 mg/L) at 72 and 96 h. According to the study, Fast Pyrolysis Bio-oil showed only slight toxicity to Danio rerio, as the determined LC₅₀ (96 h) value was between 10 and 100 mg/L (96 h LC50_zebrafish = 63.7 mg/L, analyzed concentration). No sublethal effects were observed up to and including 45.5 mg/L (nominal loading rate), and related to the analyzed concentrations, the NOEC (96 h) mortality was at 28.8 mg/L.

 

The study is reliable without restrictions.

Description of key information

Short-term toxicity of the test material (Fast Pyrolysis Bio-oil, WAF) to zebrafish (Danio rerio) was determined a in GLP-compliant study according to OECD Guidelines for Testing Chemicals (1992) no 203 "Fish, acute toxicity tests". Mortality was observed at the two highest loading rates (45.5 and 100 mg/L) at 72 and 96 h, and LC50 was determined at 63.7 mg/L (analyzed concentration).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

63.7 mg/L

Additional information

Due to poor water solubility of the substance, the organisms were exposed to a Water Accommodated Fraction (WAF) of the test substance. This is a commonly accepted procedure for regulatory studies on poorly water soluble substances.