Hydrocarbons, C11-C16, n-alkanes, isoalkanes,

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

The study was conducted between 04 August 2014 and 08 August 2014.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The hypothesis for the analogue approach is that the test substance, Hydrocarbons, C10-C13, n-alkanes, isoalkanes, <2% aromatics contains constituents which are also constituents of the target substance, Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. The substances have overlapping constituents and therefore have qualitatively similar properties (RAAF Scenario 2 applies).

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

Nominal amounts of test item (23 and 230 mg) were each separately added to the surface of 2.3 litres of test water to give the 10 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 10 and 100 mg/L loading rate WAFs.

Total Organic Carbon (TOC) analysis was performed on the test solutions at 0 and 48 hours.

Test organisms (species):

Daphnia magna

Details on test organisms:

The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.

Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room at approximately 20 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Nominal and measured concentrations:

Nominal: 10 and 100 mg/L

Details on test conditions:

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the study the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item.

Previous studies conducted on similar test items (e.g. Harlan Study Number 41304080) indicated that a 23-Hour stirring period followed by a 1-Hour standing period was sufficient to ensure that the maximum dissolved test item concentration was obtained in a Water Accommodated Fraction (WAF), as no significant increase in dissolved carbon was obtained by extending the stirring period to 96 hours.

Definitive Test
The following loading rates were assigned to the definitive test: 10 and 100 mg/L.


Experimental Preparation
Nominal amounts of test item (23 and 230 mg) were each separately added to the surface of
2.3 liters of test water to give the 10 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 10 and 100 mg/L loading rate WAFs.

Total Organic Carbon (TOC) analysis was performed on the test solutions at 0 and 48 hours.


Exposure Conditions
In the definitive test 100 mL nominal volume conical flasks containing approximately 130 mL of test preparation were used. At the start of the test 5 daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. The test vessels were then sealed to reduce loss through volitilization and maintained in a temperature controlled room at approximately 21 °C with a photoperiod of 16 hours light (398 to 481 lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.

The control group was maintained under identical conditions but not exposed to the test item.

The test preparations were not renewed during the exposure period.

Data from the control group was shared with similar concurrent studies.
Evaluations
Test Organism Observations
Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that Daphnia were considered to be immobilized if they were unable to swim for approximately 15 seconds after gentle agitation.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: The toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole.

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Total Organic Carbon Analysis
Total Organic Carbon (TOC) analysis of the test preparations at 0 and 48 hours showed no significant differences in the amount of carbon present within the 10 and 100 mg/L loading rate WAF test vessels when compared to the control vessels. Therefore, given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

The dissolved test item may have been one or several components of the test item. Given that toxicity cannot be attributed to a single component or mixture of components but to the test item as a whole, the results were based on nominal loading rates only.


Immobilization Data
There was no significant immobilization in 20 daphnids exposed to concentrations of 10 and 100 mg/L loading rate WAF for a period of 48 hours. Inspection of the immobilization data gave the following results:
The 24 h EL50 was > 100 mg/L Loading Rate WAF
The 48 h EL50 was > 100 mg/L Loading Rate WAF

The No Observed Effect Loading rate after 24 and 48 hours exposure was 100 mg/L loading rate WAF.


Validation Criteria
The test was considered to be valid given that none of the control daphnids showed immobilization or other signs of disease or stress and that the oxygen concentration at the end of the test was ≥3 mg/L in the control and test vessels.

Water Quality Criteria
Temperature was maintained at approximately 21 °C throughout the test, while there were no treatment related differences for oxygen concentration or pH.


Vortex Depth Measurements
The vortex depth was recorded at the start and end of the mixing period and was observed to be a dimple at the water surface on each occasion.


Observations on Test Item Solubility
Observations on the test media were carried out during the mixing and testing of the WAFs.

At the start of the mixing period the 10 and 100 mg/L loading rates were observed to be clear colorless water columns with globules of test item at the surface. After 23 hours stirring the 10 and 100 mg/L loading rates were observed to be clear colorless water columns. After a 1-Hour settlement period, the 10 and 100 mg/L loading rates were observed to be clear colorless solutions. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 10 and 100 mg/L loading rates were observed to be clear, colorless solutions.

Results with reference substance (positive control):

A positive control (Harlan Study Number 41400711) used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.

Exposure conditions for the positive control were similar to those in the definitive test.

Analysis of the immobilization data by the maximum-likelihood probit method (Finney, 1971*) at 24 hours and by the trimmed Spearman-Karber method (Hamilton et al 1977**) at 48 hours based on the nominal test concentrations gave the following results:
The 24 h EC50 was 0.87 mg/L with 95% confidence limits of 0.75 - 1.0 mg/L.
The 48 h EC50 was 0.71 mg/L with 95 % confidence limits of 0.65 - 0.78 mg/L.
The NOEC was 0.56 mg/L and the LOEC was 1.0 mg/L.

The No Observed Effect Concentration is based upon less than 10% immobilization at this concentration.

The results from the positive control with potassium dichromate were within the normal range for this reference item.

Reported statistics and error estimates:

An estimate of the EL50 values was given by inspection of the immobilization data.

Validation Criteria

The results of the test are considered valid if the following performance criteria are met:

·        No more than 10% of the controldaphnids show immobilization or other signs of disease or stress (e.g. discoloration or unusual behavior such as trapping at the surface water).

·        The dissolved oxygen concentration at the end of the test is ≥3 mg/L in the control and test vessels.

Validity criteria fulfilled:

yes

Conclusions:

Measured acute (short-term) toxicity data are available for Shell GTL Solvent GS190 (Hydrocarbons, C10-C13, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna. The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50 >100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

 

Methods…….

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

 

In the definitive test, twenty daphnids (4 replicates of 5 animals) were exposed to Water Accommodated Fractions (WAFs) of the test item at nominal loading rates of 10 and 100 mg/L for 48 hours at a temperature of approximately 21°C under static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

 

Results…….

Samples of the control and the 10 and 100 mg/L loading rate WAFs were taken at 0 (fresh media) and 48 hours (old media) for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

 

Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

 

Exposure of Daphnia magna to the test item gave EL₅₀values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

The study was conducted between 04 August and 08 August.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP guideline study.

Justification for type of information:

The hypothesis for the analogue approach is that the test substance, Hydrocarbons, C12-C15, n-alkanes, isoalkanes <2% aromatics contains constituents which are also constituents of the target substance, Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. The substances have overlapping constituents and therefore have qualitatively similar properties (RAAF Scenario 2 applies).

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.

Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room at approximately 20 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Nominal and measured concentrations:

Nominal: 10 and 100 mg/L.

Details on test conditions:

Procedure
Due to the low aqueous solubility and complex nature of the test item, for the purposes of the study the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item.

Previous studies conducted on similar test items (e.g. Harlan Study Number 41304080) indicated that a 23-Hour stirring period followed by a 1-Hour standing period was sufficient to ensure that the maximum dissolved test item concentration was obtained in a Water Accommodated Fraction (WAF), as no significant increase in dissolved carbon was obtained by extending the stirring period to 96 hours.


Definitive Test
The following loading rates were assigned to the definitive test: 10 and 100 mg/L.


Experimental Preparation
Nominal amounts of test item (23 and 230 mg) were each separately added to the surface of
2.3 liters of test water to give the 10 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 10 and 100 mg/L loading rate WAFs.

Total Organic Carbon (TOC) analysis was performed on the test solutions at 0 and 48 hours.


Exposure Conditions
In the definitive test 100 mL nominal volume conical flasks containing approximately 130 mL of test preparation were used. At the start of the test 5 daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. The test vessels were then sealed to reduce loss through volatilization and maintained in a temperature controlled room at approximately 21 °C with a photoperiod of 16 hours light (398 to 481 lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.

The control group was maintained under identical conditions but not exposed to the test item.
The test preparations were not renewed during the exposure period.
Data from the control group was shared with similar concurrent studies.


Evaluations
Test Organism Observations
Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that Daphnia were considered to be immobilized if they were unable to swim for approximately 15 seconds after gentle agitation.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: The toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole.

Details on results:

Total Organic Carbon Analysis
Total Organic Carbon (TOC) analysis of the test preparations at 0 and 48 hours showed no significant differences in the amount of carbon present within the 10 and 100 mg/L loading rate WAF test vessels when compared to the control vessels. Therefore, given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

The dissolved test item may have been one or several components of the test item. Given that toxicity cannot be attributed to a single component or mixture of components but to the test item as a whole, the results were based on nominal loading rates only.


Immobilization Data
There was no immobilization in 20 daphnids exposed to concentrations of 10 and 100 mg/L loading rate WAF for a period of 48 hours. Inspection of the immobilization data gave the following results:
The 24 h EL50 was > 100 mg/L Loading Rate WAF
The 48 h EL50 was > 100 mg/L Loading Rate WAF

The No Observed Effect Loading rate after 24 and 48 hours exposure was 100 mg/L loading rate WAF.

Results with reference substance (positive control):

A positive control (Harlan Study Number 41400711) used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.
Exposure conditions for the positive control were similar to those in the definitive test.

Analysis of the immobilization data by the maximum-likelihood probit method (Finney, 1971*) at 24 hours and by the trimmed Spearman-Karber method (Hamilton et al 1977**) at 48 hours based on the nominal test concentrations gave the following results:

The 24 h EC50 was 0.87 mg/L with 95 % confidence limits of 0.75 – 1.0 mg/L.
The 48 h EC50 was 0.71 mg/L with 95 % confidence limits of 0.65 – 0.78 mg/L.
The NOEC was 0.56 mg/L and the LOEC was 1.0 mg/L.

The No Observed Effect Concentration is based upon less than 10% immobilization at this concentration.
The results from the positive control with potassium dichromate were within the normal range for this reference item.

Reported statistics and error estimates:

An estimate of the EL50 values was given by inspection of the immobilization data.

Validation Criteria

The test was considered to be valid given that none of the control daphnids showed immobilization or other signs of disease or stress and that the oxygen concentration at the end of the test was ≥3 mg/L in the control and test vessels.

 

Water Quality Criteria

Temperature was maintained at approximately 21°C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

 

Vortex Depth Measurements

The vortex depth was recorded at the start and end of the mixing period and was observed to be a dimple at the water surface on each occasion.

 

Observations on Test Item Solubility

Observations on the test media were carried out during the mixing and testing of the WAFs.

 

At the start of the mixing period the 10 and 100 mg/L loading rates were observed to be clear colorless water columns with globules of test item at the surface. After 23 hours stirring and the 1-Hour settlement period, the 10 and 100 mg/L loading rates were observed to be clear colorless solutions. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 10 and 100 mg/L loading rates were observed to be clear, colorless solutions. 

 

Validity criteria fulfilled:

yes

Conclusions:

Measured acute (short-term) toxicity data are available for Shell GTL Solvent G215 (Hydrocarbons, C12-C15, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna. The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50 >100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

 

Methods…….

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

 

In the definitive test, twenty daphnids (4 replicates of 5 animals) were exposed to Water Accommodated Fractions (WAFs) of the test item at nominal loading rates of 10 and 100 mg/L for 48 hours at a temperature of approximately 21°C under static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

 

Results…….

Samples of the control and the 10 and 100 mg/L loading rate WAFs were taken at 0 (fresh media) and 48 hours (old media) for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

 

Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

 

Exposure of Daphnia magna to the test item gave EL₅₀values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

The study was conducted between 05 August 2014 and 13 August 2014.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP guideline study.

Justification for type of information:

The hypothesis for the analogue approach is that the test substance, Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics contains constituents which are also constituents of the target substance, Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. The substances have overlapping constituents and therefore have qualitatively similar properties (RAAF Scenario 2 applies).

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.

Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room at approximately 20 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Nominal and measured concentrations:

Nominal: 10 and 100 mg/L.

Details on test conditions:

Procedure
Due to the low aqueous solubility and complex nature of the test item, for the purposes of the study the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item.

Previous studies conducted on similar test items (e.g. Harlan Study Number 41304080) indicated that a 23-Hour stirring period followed by a 1-Hour standing period was sufficient to ensure that the maximum dissolved test item concentration was obtained in a Water Accommodated Fraction (WAF), as no significant increase in dissolved carbon was obtained by extending the stirring period to 96 hours..


Definitive Test
The following loading rates were assigned to the definitive test: 10 and 100 mg/L.

Experimental Preparation
Nominal amounts of test item (23 and 230 mg) were each separately added to the surface of
2.3 litres of test water to give the 10 and 100 mg/L loading rates respectively. Due to the volatile nature of the test item, all stirring vessels were completely filled and sealed. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 10 and 100 mg/L loading rate WAFs.

Total Organic Carbon (TOC) analysis was performed on the test solutions at 0 and 48 hours.

Exposure Conditions
In the definitive test completely filled 100 mL nominal volume conical flasks containing approximately 130 mL of test preparation were used. At the start of the test 5 daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. The test vessels were then sealed to reduce loss through volitilization and maintained in a temperature controlled room at approximately 21 °C with a photoperiod of 16 hours light (618 to 645 lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.

The control group was maintained under identical conditions but not exposed to the test item.

The test preparations were not renewed during the exposure period.

Control data was shared with similar concurrent studies.


Evaluations
Test Organism Observations
Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that Daphnia were considered to be immobilized if they were unable to swim for approximately 15 seconds after gentle agitation.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: The toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole.

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Total Organic Carbon Analysis
Total Organic Carbon (TOC) analysis of the test preparations at 0 and 48 hours showed no significant differences in the amount of carbon present within the 10 and 100 mg/L loading rate WAF test vessels when compared to the control vessels. Therefore, given the low background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

The dissolved test item may have been one or several components of the test item. Given that toxicity cannot be attributed to a single component or mixture of components but to the test item as a whole, the results were based on nominal loading rates only.

Immobilization Data
There was no immobilization in 20 daphnids exposed to concentrations of 10 and 100 mg/L loading rate WAF for a period of 48 hours. Inspection of the immobilization data gave the following results:
The 24 h EL50 was > 100 mg/L Loading Rate WAF
The 48 h EL50 was > 100 mg/L Loading Rate WAF

The No Observed Effect Loading rate after 24 and 48 hours exposure was 100 mg/L loading rate WAF.

Results with reference substance (positive control):

A positive control (Harlan Study Number 41400711) used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.
Exposure conditions for the positive control were similar to those in the definitive test.

Analysis of the immobilization data by the maximum-likelihood probit method (Finney, 1971 ) at 24 hours and by the trimmed Spearman-Karber method (Hamilton et al 1977**) at 48 hours based on the nominal test concentrations gave the following results:
The 24 h EC50 was 0.87 mg/L wth 95 % confidence limits of 0.75 – 1.0 mg/L.
The 48h EC50 was 0.71 mg/L wth 95 % confidence limits of 0.65 – 0.78 mg/L.
The NOEC was 0.56 mg/L and the LOEC was 1.0 mg/L.

The No Observed Effect Concentration is based upon less than 10% immobilization at this concentration.
The results from the positive control with potassium dichromate were within the normal range for this reference item.

Reported statistics and error estimates:

An estimate of the EL50 values was given by inspection of the immobilization data.

Validation Criteria

The test was considered to be valid given that none of the control daphnids showed immobilization or other signs of disease or stress and that the oxygen concentration at the end of the test was ≥3 mg/L in the control and test vessels.

 

Water Quality Criteria

Temperature was maintained at approximately 20°C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

 

Vortex Depth Measurements

The vortex depth was recorded at the start and end of the mixing period and was observed to be a dimple at the water surface on each occasion.

 

Observations on Test Item Solubility

Observations on the test media were carried out during the mixing and testing of the WAFs.

 

At the start of the mixing period the 10 and 100 mg/L loading rates were observed to be clear colorless water columns with globules of test item at the surface. After 23 hours stirring and a
1-Hour standing period the 10 and 100 mg/L loading rates were observed to be clear colorless water columns with a slick of test item on the surface. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 10 and 100 mg/L loading rates were observed to be clear, colorless solutions. 

Validity criteria fulfilled:

yes

Conclusions:

Measured acute (short-term) toxicity data are available for Shell GTL Solvent GS250 (Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna. The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50 >100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

 

Methods…….

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

 

In the definitive test, twenty daphnids (4 replicates of 5 animals) were exposed to Water Accommodated Fractions (WAFs) of the test item at nominal loading rates of 10 and 100 mg/L for 48 hours at a temperature of approximately 20°C under static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

 

Results…….

Samples of the control and the 10 and 100 mg/L loading rate WAFs were taken at 0 (fresh media) and 48 hours (old media) for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

 

Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

 

Exposure of Daphnia magna to the test item gave EL₅₀values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

The study was conducted between 05 August 2014 and 13 August 2014.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Justification for type of information:

The hypothesis for the analogue approach is that the test substance, Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics contains constituents which are also constituents of the target substance, Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. The substances have overlapping constituents and therefore have qualitatively similar properties (RAAF Scenario 2 applies).

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.

Adult Daphnia were maintained in 150 mL glass beakers containing Elendt M7 medium in a temperature controlled room at approximately 20 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Nominal and measured concentrations:

Nominal: 10 and 100 mg/L

Details on test conditions:

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the study the test medium was prepared as a Water Accommodated Fraction (WAF) of the test item.

Previous studies conducted on similar test items (e.g. Harlan Study Number 41304080) indicated that a 23-Hour stirring period followed by a 1-Hour standing period was sufficient to ensure that the maximum dissolved test item concentration was obtained in a Water Accommodated Fraction (WAF), as no significant increase in dissolved carbon was obtained by extending the stirring period to 96 hours.


Definitive Test
The following loading rates were assigned to the definitive test: 10 and 100 mg/L.


Experimental Preparation
Nominal amounts of test item (23 and 230 mg) were each separately added to the surface of
2.3 litres of test water to give the 10 and 100 mg/L loading rates respectively. Due to the volatile nature of the test item, all stirring vessels were completely filled and sealed. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid-depth siphoning (the first approximate 75-100 mL discarded) to give the 10 and 100 mg/L loading rate WAFs.

Total Organic Carbon (TOC) analysis was performed on the test solutions at 0 and 48 hours.


Exposure Conditions
In the definitive test completely filled 100 mL nominal volume conical flasks containing approximately 130 mL of test preparation were used. At the start of the test 5 daphnids were placed in each test and control vessel at random, in the test preparations. Four replicate test and control vessels were prepared. The test vessels were then sealed to reduce loss through volitilization and maintained in a temperature controlled room at approximately 21 °C with a photoperiod of 16 hours light (618 to 645 lux) and 8 hours darkness with 20 minute dawn and dusk transition periods. The daphnids were not individually identified, received no food during exposure and the test vessels were not aerated.

The control group was maintained under identical conditions but not exposed to the test item.

The test preparations were not renewed during the exposure period.

Control data was shared with similar concurrent studies.


Evaluations
Test Organism Observations
Any immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure. The criterion of effect used was that Daphnia were considered to be immobilized if they were unable to swim for approximately 15 seconds after gentle agitation.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: The toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole.

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Total Organic Carbon Analysis
Total Organic Carbon (TOC) analysis of the test preparations at 0 and 48 hours showed no significant differences in the amount of carbon present within the 10 and 100 mg/L loading rate WAF test vessels when compared to the control vessels. Therefore, given the low background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

The dissolved test item may have been one or several components of the test item. Given that toxicity cannot be attributed to a single component or mixture of components but to the test item as a whole, the results were based on nominal loading rates only.


Immobilization Data
There was no immobilization in 20 daphnids exposed to concentrations of 10 and 100 mg/L loading rate WAF for a period of 48 hours. Inspection of the immobilization data gave the following results:

The 24 h EL50 was > 100 mg/L Loading Rate WAF
The 48 h EL50 was > 100 mg/L Loading Rate WAF

The No Observed Effect Loading rate after 24 and 48 hours exposure was 100 mg/L loading rate WAF.

Results with reference substance (positive control):

A positive control (Harlan Study Number 41400711) used potassium dichromate as the reference item at concentrations of 0.32, 0.56, 1.0, 1.8 and 3.2 mg/L.

Exposure conditions for the positive control were similar to those in the definitive test.

Analysis of the immobilization data by the maximum-likelihood probit method (Finney, 1971 ) at 24 hours and by the trimmed Spearman-Karber method (Hamilton et al 1977**) at 48 hours based on the nominal test concentrations gave the following results:
The 24 h EC50 was 0.87 mg/L with 95 % confidence limits of 0.75 – 1.0 mg/L.
The 48 h EC50 was 0.71 mg/L with 95 % confidence limits of 0.65 – 0.78 mg/L.
The NOEC was 0.56 mg/L and the LOEC was 1.0 mg/L.

The No Observed Effect Concentration is based upon less than 10% immobilization at this concentration.

The results from the positive control with potassium dichromate were within the normal range for this reference item.

Reported statistics and error estimates:

An estimate of the EL50 values was given by inspection of the immobilization data.

Validation Criteria

The test was considered to be valid given that none of the control daphnids showed immobilization or other signs of disease or stress and that the oxygen concentration at the end of the test was ≥3 mg/L in the control and test vessels.

 

Water Quality Criteria

Temperature was maintained at approximately 20°C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

 

Vortex Depth Measurements

The vortex depth was recorded at the start and end of the mixing period and was observed to be a dimple at the water surface on each occasion.

 

Observations on Test Item Solubility

Observations on the test media were carried out during the mixing and testing of the WAFs.

At the start of the mixing period the 10 and 100 mg/L loading rates were observed to be clear colorless water columns with globules of test item at the surface. After 23 hours stirring and a
1-Hour standing period the 10 and 100 mg/L loading rates were observed to be clear colorless water columns with a slick of test item on the surface. Microscopic inspection of the WAFs showed no micro-dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 10 and 100 mg/L loading rates were observed to be clear, colorless solutions. 

Validity criteria fulfilled:

yes

Conclusions:

Measured acute (short-term) toxicity data are available for Shell GTL Solvent GS270 (Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna. The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50 >100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphnia sp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

 

Methods…….

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

 

In the definitive test, twenty daphnids (4 replicates of 5 animals) were exposed to Water Accommodated Fractions (WAFs) of the test item at nominal loading rates of 10 and 100 mg/L for 48 hours at a temperature of approximately 20°C under static test conditions. The number of immobilized Daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

 

Results…….

Samples of the control and the 10 and 100 mg/L loading rate WAFs were taken at 0 (fresh media) and 48 hours (old media) for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method.

 

Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

 

Exposure of Daphnia magna to the test item gave EL₅₀values of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

 

Description of key information

There is no data available for Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. However, data is available for structural analogues, Hydrocarbons, C10-C13, n-alkanes, isoalkanes, <2% aromatics; Hydrocarbons, C12-C15, n-alkanes, isoalkanes, <2% aromatics; and Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics and presented in the dossier. The hypothesis for the analogue approach is that the test substances, contain constituents which are also constituents of the target substance, Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. The substances have overlapping constituents and therefore have qualitatively similar properties (RAAF Scenario 2 applies).

Key information is summarised below:

Water accommodated fractions of Hydrocarbons, C10-C13, n-alkanes, isoalkanes, <2% aromatics, produced a 48-hour EL50 value of >100 mg/L in Daphnia magna. No mortality was observed in either the control or treatment test systems.

Water accommodated fractions of Hydrocarbons, C12-C15, n-alkanes, isoalkanes, <2% aromatics, produced a 48-hour EL50 value of >100 mg/L in Daphnia magna. No mortality was observed in either the control or treatment test systems.

Water accommodated fractions of Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics, produced a 48-hour EL50 value of >100 mg/L in Daphnia magna. No mortality was observed in either the control or treatment test systems. The 48h-LL50 is > 100 mg/l.

Water accommodated fractions of Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics, produced a 48-hour LL50 value of >100 mg/L in Daphnia magna.

Additional information

There is no data available for Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. However, data is available for structural analogues, Hydrocarbons, C10-C13, n-alkanes, isoalkanes, <2% aromatics; Hydrocarbons, C12-C15, n-alkanes, isoalkanes, <2% aromatics; and Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics and presented in the dossier. The hypothesis for the analogue approach is that the test substances, contain constituents which are also constituents of the target substance, Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics. The substances have overlapping constituents and therefore have qualitatively similar properties (RAAF Scenario 2 applies).

No measured short-term toxicity to aquatic invertebrate data are available for the registration substance. However, reliable data are available for closely related substances covering the same carbon number range, including other Fischer-Tropsch process-derived materials.

 

Measured acute (short-term) toxicity data are available for Shell GTL Solvent GS190 (Hydrocarbons, C10-C13, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna (Shell, 2014). The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50>100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Measured acute (short-term) toxicity data are available for Shell GTL Solvent G215 (Hydrocarbons, C12-C15, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna (Shell, 2014). The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50>100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Measured acute (short-term) toxicity data are available for Shell GTL Solvent GS250 (Hydrocarbons, C14-C16, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna (Shell, 2014). The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50>100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

Measured acute (short-term) toxicity data are available for Shell GTL Solvent GS270 (Hydrocarbons, C15-C19, n-alkanes, isoalkanes, <2% aromatics) to the freshwater invertebrate Daphnia magna (Shell, 2014). The test was conducted under static (no renewal of the test media) conditions in accordance with OECD Test Guideline 202 and method C2 of EC Regulation No. EC 440/2008. Appropriate modifications to the test and media preparation procedures were made to take account of the test substance containing multiple constituents, having low solubility in water and being potentially volatile. No effects on mobility of D. magna were observed after 48 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates up to 100 mg/l; 48-hour EL50>100 mg/l, NOELR ≥100 mg/l. Samples of the control and the 10 and 100 mg/l loading rate WAFs were taken at 0 and 48 hours for Total Organic Carbon (TOC) analysis. Given the background level of carbon in the control vessels and also the low level of carbon in the test vessels, it was considered that all the results were around the limit of quantification of the analytical method. The results of the test are considered to be reliable.

These data are used as weight of evidence to demonstrate that Hydrocarbons, C11-C16, n-alkanes, isoalkanes, <2% aromatics is not acutely harmful to aquatic invertebrates.