Hydrocarbons, C18-C24, iso-alkanes <2% aromatics

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:

04 August 2014 to 08 August 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

This study has been performed according to OECD and/or EC guidelines and according to GLP principles

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 2: Semi-static method)

Version / remarks:

1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material:
Hygroscopic: Not indicated
Volatile: Not indicated
Specific Gravity / Density: <800 kg/m3 at 15 C
Stability in water: Not indicated
Solubility in water: Insoluble

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all
- Sampling method: Approximately 50 ml from the approximate centre of the test vessels. The sample vessels were completely filled and closed airtight.
- Sample storage conditions before analysis: Samples were stored in a refrigerator until analysis.

Vehicle:

no

Details on test solutions:

Preparation was performed in closed vessels with a tap. The headspace was reduced to a minimum. Individual loading rates of 1.0, 10 and 100 mg/l were prepared. A 1-day period of magnetic stirring was applied. During the stirring period care was taken to prevent the formation of an emulsion. Therefore, the vortex was in all cases less than 10% of the height of the medium, preventing the test substance being pulled down and hitting the stirring bar. The vortex was similar in all vessels. At the end of the stirring a floating layer and droplets of the test substance suspended in the solution were observed in the vessels. After a 1-hour settlement period the aqueous WAFs were collected using a tap at the bottom of the vessel and used as test concentrations (a small volume of the WAF used to flush the tap was discarded). A macroscopic observation of the final test solutions revealed that they were all colorless and free of undissolved material.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Length at study initiation (mean and SD): 2.2 ± 0.1 cm
- Weight at study initiation (mean and SD): 0.22 ± 0.05 g
- Method of breeding: F1 from a single parent-pair bred in UV-treated water.
- Feeding during test: no

ACCLIMATION
- Acclimation period: Fish were hold in a holding tank with comparable water quality parameters and pH and temperature differences between test and holding tank media of less than 0.5 unit and 0.5°C. Therefore, no acclimation was needed.

QUARANTINE (wild caught)
- Duration: at least 12 days
- Health/mortality: less than 5% during the seven days prior to the start of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

no

Hardness:

180 mg/l expressed as CaCO3

Test temperature:

22-23 C

pH:

7.5-7.9

Dissolved oxygen:

6.3-9.1

Salinity:

not measured

Nominal and measured concentrations:

control and a Water Accommodated Fraction (WAF) prepared at a loading rate of 100 mg/l (limit test); in a combined range-finding test WAFs prepared at 1.0 and 10 mg/l were tested.

TOC-concentrations at the WAF prepared at 100 mg/l could not be reliably determined as they were below the LOD (~1 mg TOC/l) of the equipment.

Details on test conditions:

TEST SYSTEM
- Test vessel: 3.5 L
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: all glass, 3.3 L test solution
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7 (limit test); 3 per vessel for loading rates of 1.0 and 10 mg/l (combined range-finding test)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 0 - no vehicle control
- Biomass loading rate:
For the control and the WAF prepared at 100 mg/l: 0.47 g fish/litre, i.e. 7 fish per 3.3 litres of test medium,
For the WAFs prepared at 1.0 and 10 mg/l: 0.20 g fish/litre, i.e. 3 fish per 3.3 litres of test medium.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: adjusted ISO medium

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : 3.25, 24, 48, 72 and 96h LL50

TEST CONCENTRATIONS
- Range finding study: The study was performed as a combined limit/range-finding test

Reference substance (positive control):

yes

Remarks:

pentachlorophenol

Duration:

3.25 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Water-accommodated fraction

Duration:

24 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Water-accommodated fraction

Duration:

48 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Water-accommodated fraction

Duration:

72 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Water-accommodated fraction

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Water-accommodated fraction

Details on results:

- Behavioural abnormalities: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- LC50: 0.24 mg/l

Reported statistics and error estimates:

No LL50 could be determined because the test substance proved to be non-toxic (LL50 > maximum soluble concentration).

Validity criteria fulfilled:

yes

Conclusions:

Measured acute (short-term) toxicity data are available for Hydrocarbons, C18-C24, isoalkanes, <2% aromatics to the freshwater fish Cyprinus carpio (carp). The test was conducted under semi-static conditions (daily renewal of the test media) in accordance with OECD Test Guideline 203. 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 in accordance with OECD guidance on aquatic toxicity testing of difficult substances and mixtures (OECD, 2000). No mortality or clinical effects in C. carpio were observed after 96 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates of up to 100 mg/l; 96-hour LL50 >100 mg/l, NOELR ≥100 mg/l. TOC concentrations were analysed in samples taken from the control and WAF prepared at 100 mg/l. In all cases the concentrations were below the limit of detection of the method (1.0 mg/l). The results of the test are considered to be reliable.

Description of key information

A 96-hour LL₅₀ value of >100 mg/l and NOELR value of ≥100 mg/l (WAF loading rate) (highest concentration tested) have been determined for the effects of the test substance on mortality and sublethal effects of Cyprinus carpio.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

Measured acute (short-term) toxicity data are available for Hydrocarbons, C18-C24, isoalkanes, <2% aromatics to the freshwater fish Cyprinus carpio (carp) (Tobor-Kaplan, 2014). The test was conducted under semi-static conditions (daily renewal of the test media) in accordance with OECD Test Guideline 203. 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 in accordance with OECD guidance on aquatic toxicity testing of difficult substances and mixtures (OECD, 2000). No mortality or clinical effects in C. carpio were observed after 96 hours exposure to the test medium prepared as a water-accommodated fraction (WAF) at loading rates of up to 100 mg/l; 96-hour LL50 >100 mg/l, NOELR ≥100 mg/l. TOC concentrations were analysed in samples taken from the control and WAF prepared at 100 mg/l. In all cases the concentrations were below the limit of detection of the method (1.0 mg/l). The results of the test are considered to be reliable.

The acute toxicity of a sample of GTL Gasoil (Shell SMDS-2) was determined by Palmer (2002) in a test with the fish Pimephales promelas (Fathead minnow). The test was conducted in accordance with OECD Test Guideline 203.

Water-accommodated fractions (WAFs) of the sample were prepared in sealed vessels with minimum headspace, by stirring for approximately 72 hours. The contents of the vessels were left to stand for 1-2 hours before drawing off the aqueous phase – the WAF – for testing. Semi-static exposures were then carried out in completely full, sealed vessels. The tests were not subject to GLP and the test media were not analysed for stability or for exposure concentration of the test substance. However in other respects they were considered to fulfil the requirements of current best practice.

The test results, expressed as the LL₅₀ value, showed that the sample was not acutely toxic to fish at a loading rate of 1000 mg/l.

Results have been reported for an acute toxicity test carried out on a second sample of the substance (SASOL SPD™ Diesel) with the fish Brachydanio rerio (Zebrafish) (Albertus and Phillips, 2005). The test was conducted in accordance with OECD Test Guideline 203, but was not subject to GLP and no analysis of test media was reported. The test media were WAFs of the test sample that were prepared in sealed vessels with a small headspace according to the CONCAWE methodology for the ecotoxicological testing of petroleum products (CONCAWE report, 1993).

The test results, expressed as the LL₅₀ value, showed that the sample was not acutely toxic to fish at a loading rate of 1000 mg/l.

The short-term toxicity of GTL Base Oil Distillates to freshwater fish Brachydanio rerio (zebra fish) has been determined by Han (2008). The test was conducted in accordance with OECD Test Guideline 203.

Water-accommodated fractions (WAFs) of the sample were prepared in jars, by stirring for approximately 24 hours. The contents of the vessels were left to stand for 2 hours before drawing off the aqueous phase – the WAF – for testing. Semi-static exposures were then carried out.

The test was conducted in accordance with GLP and the TOC of test media were analysed for stability and exposure concentration. Total Organic Carbon (TOC) analysis of the freshly prepared test preparations showed amount of carbon present within the 1000 mg/l loading rate WAF test vessels to range from 1.71 mg C/l to 1.79 mg C/l. TOC analysis of the old test media showed amount of carbon present to range from 1.56 mg C/l to 1.64 mg C/l. There was no significant difference in TOC content between control and exposure vessels. This was expected by the authors due to the low water solubility of the test substance. 

The test results, expressed as the LL₅₀ value, showed that GTL Base Oil Distillates was not hazardous to fish at a loading rate of 1000 mg/l WAFs.

Conclusion

Short-term fish toxicity tests are available for the registered substance. In the absence of adverse effects in this study it can be concluded that the LLR₅₀ (freshwater fish) for Hydrocarbons, C18-C24, isoalkanes, <2% aromatics is >100 mg/l.

Reference

Concawe 1993 Ecotoxicological testing of petroleum products: test methodology. Report no. 92/56. 21 Jan 1993.