Fast Pyrolysis Bio-oil

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 June 2013 - 16 July 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

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

Analytical monitoring:

yes

Details on sampling:

Samples from fresh and aged test solutions were taken at three representative sampling intervals from one renewal period of each week and at the end of the test. All concentration levels and the control were analysed on days 0, 9 and 16 for fresh samples and days 2, 12 and 19 for aged samples.

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 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 The test solution volume of 50 mL per test vessel was withdrawn from the middle of the preparation vessel by suction (according to OECD SERIES ON TESTING AND ASSESSMENT No. 23). Transfer of undissolved test item was avoided. All solutions were prepared freshly prior to each day of application/renewal.

Test organisms (species):

Daphnia magna

Details on test organisms:

As test organism, Daphnia magna STRAUS, Clone V, was used. The animals are continuously bred in the laboratory according to the standard operating procedures. The Daphnia were originally purchased in a healthy condition from the Federal Environmental Agency in Berlin/Germany.

Test type:

semi-static

Water media type:

not specified

Limit test:

no

Total exposure duration:

21 d

Hardness:

total hardness was between 13 and 14

Test temperature:

18.8 - 21.0°C

pH:

6.81 - 8.69

Dissolved oxygen:

> 71%

Salinity:

not determined

Nominal and measured concentrations:

The results were evaluated in a semi-static test with nominal concentrations of 0, 2.56, 6.40, 16.0, 40.0 and 100 mg/L with renewal of test solutions every Monday, Wednesday and Friday.
In the fresh test solutions the mean measured content of Fast Pyrolysis Bio-oil was 50 % of the nominal concentration. In the aged test solutions the mean measured content of Fast Pyrolysis Bio-oil was 12 % of nominal.
The content of Fast Pyrolysis Bio-oil in the fresh samples of the highest test item concentration of 100 mg/L was between 85 % and 89 %. The time weighted mean of the measured concentrations was calculated.

Details on test conditions:

The test was performed with 10 vessels for each test item concentration and the control, containing 1 Daphnia each. The reproduction test was performed semi-statically with renewal of test solutions 3 times a week (every Monday, Wednesday and Friday).

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

39.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 39.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

39.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 39.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

> 39.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 39.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

No Mortality of adult Daphnia above the allowed control mortality was observed in the control and all test item concentrations.
The first offspring in the test item concentrations of 2.56, 6.40 and 16.0 mg/L was observed on day 9, at 40.0 mg/L on day 10 and in the control and 100 mg/L on day 11.
No toxic effects were observed up to 100 mg/L (nominal).

Results with reference substance (positive control):

not available

Reported statistics and error estimates:

No Mortality of adult Daphnia above the allowed control mortality was observed in the control and all test item concentrations. Also there was no impact of the test item on the reproductive capacity or the mortality of offspring observed. Therefore no statistical evaluation was indicated for those parameters.

Table10:      Number of living offspring (0 d - 21 d) per adult (alive at the end of the test and inhibition of reproduction (%)

- = not applicable

Concentration [mg/L]	Mean number of alive offspring per adult	Inhibition [%]
Control	78.4	-
2.56	94.2	- 20.2
6.40	125.7	- 60.3
16.0	148	- 88.8
40.0	113	- 44.1
100	72.7	7.3

Validity criteria fulfilled:

yes

Conclusions:

Long-term toxicity of fast pyrolysis bio-oil (water accommodated fraction) to Daphnia magna was determined in a D. magna reproduction test. No statistically significant adverse effects were observed in the test. EC50 was determined to be above the highest loading rate for both, adult mortality and effects on reproduction (EC50, 21 d, nominal > 100 mg/L and EC50, 21 d, analyzed concentration = 37.9 mg/L).

Executive summary:

Long-term toxicity of fast pyrolysis bio-oil (tested as water accommodated fraction) to water flea Daphnia magna was determined in a GLP compliant study according to OECD Guideline no 211 (2012) "Daphnia magna Reproduction test". Five nominal concentrations in the range between 2.56 and 100 mg/L differing by a dilution factor of 2.5 were tested. The control group was exposed to untreated test medium.

 

The following findings can be summarised from the tables presented above:

The EC50 for inhibition of reproduction is assumed to be > 100 mg/L.

The NOEC (21 d) for inhibition of reproduction was determined to be 100 mg/L. The LOEC (21 d) for inhibition of reproduction is estimated to be > 100 mg/L.

The LC50 (21d) is assumed to be > 100 mg/L. The NOEC (21 d) for mortality of adults was observed at 100 mg/L. The LOEC (21 d) for mortality of adults is estimated to be > 100 mg/L.

The mortality of parent animals (female Daphnia) did not exceed 20% at the end of the test in the control. The mean number of living offspring produced per parent animal alive at the end of the test was ≥ 60 in the control. The test is therefore regarded as valid.

 

Relating the concentrations analysed, the following results were obtained:

The EC50 for inhibition of reproduction (21 d) was assumed to be > 39.7 mg/L. The NOEC (21 d) for inhibition of reproduction was determined to be 39.7 mg/L. The LOEC (21 d) for inhibition of reproduction was assumed to be > 39.7 mg/L.

The LC50 (21d) was assumed to be > 39.7 mg/L. The NOEC (21 d) for mortality of adults was observed at 39.7 mg/L. The LOEC (21 d) for mortality of adults was > 39.7 mg/L.

 

According to the test, fast pyrolysis bio-oil showed low chronic toxicity to Daphnia as no statistically significant adverse effects were observed in the test: EC50 was determined to be above the highest loading rate (100 mg/L, analyzed concentration 37.9 mg/L) for both adult mortality and effects on reproduction.

 

The test is reliable without restrictions.

Description of key information

Long-term toxicity of the test material (Fast Pyrolysis Bio-oil, WAF) to water flea Daphnia magna was determined in a GLP compliant study according to OECD Guidelines for Testing Chemicals (2012) no 211 "Daphnia magna Reproduction test". No statistically significant adverse effects were observed in the test. EC50 was determined to be above the highest loading rate (100 mg/L, analyzed concentration 37.9 mg/L) for both adult mortality and effects on reproduction. 

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.