Octadecene, reaction products with hexadecene, hydrogenated

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:

09 December 2019 to 13 December 2019

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:

OECD Guideline for Testing of Chemicals, Section 2, No. 203, "Fish, Acute Toxicity Test", adopted July 17, 1992.

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Version / remarks:

EPA Guideline 712-C-96-118: OCSPP 850.1075, "Fish Acute Toxicity Test, Freshwater and Marine" April 1996

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

Commission Regulation (EU) 2017/735 of 14 February 2017 amending, for the purpose of its adaptation to technical progress, the Annex to Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), Annex IV Part C, C1.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

No further details specified in the study report.

Analytical monitoring:

yes

Details on sampling:

Duplicate samples were taken (2 x ~15 mL) in glass tubes at the beginning and at the end of the experiment from the control and at the applied test concentration level.

Vehicle:

no

Details on test solutions:

The test solutions were prepared using a saturated solution method according to the Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, OECD No. 23 (2000).
Saturated test item solution with the nominal loading rate of 100.0 mg/L (water accommodated fractions, WAF) was prepared with direct addition of the test item, mixed into the aquarium water and then continuously stirred gently for approximately 48 hours using magnetic stirrer. Then, the emulsion was settled for approximately 1 hour, and the middle layer was collected as the test solution. This test solution (100.0 mg/L WAF) was divided into two parts. The non-dissolved test material was removed by filtration through a fine (0.22 µm) filter to give the appropriate WAF solution in the first case, while the other set was not filtered and used as test solution as it is.
Prior to treatment of each renewal period, test item solutions were prepared by the method described above just before introduction of the fish (start of the treatments).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Zebrafish (Brachydanio rerio)
Source: István Szent University, 2100 Gödöllő, Páter Károly u. 1, Hungary
Justification of species: Zebrafish (Brachydanio rerio) is one of the convenient species for acute fish toxicity test.
Number of animals: seven fish per test group
Body length of animals: 2.1 – 2.6 cm
Food and Feeding: The fish were not fed during the test
Acclimatisation: more than 12 days
Animal health: Fish were bred in a well-known fish farm, under disease- and parasite-controlled conditions. Fish were observed during the acclimatisation and test period; the fish were healthy. There was no mortality of the population for seven days before the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation period specified in the study report.

Hardness:

The hardness of the dilution water was determined as 178 mg/L (as CaCO3).

Test temperature:

The test temperature was between 20.9 and 23.5 °C.

pH:

The pH was in the range of 7.83 – 8.33.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 73 – 100 % of the air saturation value at the temperature used.

Salinity:

Not specified

Conductivity:

Not specified

Nominal and measured concentrations:

nominal loading rate (WAF).

Details on test conditions:

DESCRIPTION OF THE TEST PROCEDURE
The test duration was 96 hours. One aquarium was used for test groups and for the control group, respectively. Each aquarium comprised 7 fish and 3 L test solution. The animals were not fed during the test. The loading of the test aquaria was less than 1.0 g fish/L test solution at the start of the experiment.
The test was performed under semi-static conditions. The frequency of the water renewal periods was 24 hours.
The choice of the test concentrations was made on the basis of the results of the preliminary range-finding test.

Preliminary Range-Finding Test
A preliminary concentration range-finding test was conducted to determine the approximate toxicity of the test item so that appropriate test concentrations could be selected for use in the definitive test. Four fish were exposed to each concentration of the test item plus a control, for 96 hours under semi-static conditions (with 24-hour renewal, in absence of stability data).
Saturated test item solutions with the nominal loading rates of 0.1, 1.0, 10.0 and 100.0 mg/L (water accommodated fractions, WAFs) were individually prepared with direct addition of the test item, mixed into the test medium (aquarium water) and then continuously stirred gently for approximately 48 hours using magnetic stirrer. Then, the emulsions were settled for approximately 1 hour, and the middle layer was collected as the test solution in each tested level.
Test solution at the highest test level (100.0 mg/L WAF) was divided into two parts as described above.

Test item concentrations in the Definitive Test
Because no toxic response was observed at the highest concentration level during the preliminary range-finding test, only one test concentration at 100.0 mg/L nominal loading rate (WAF) (a filtered and a non-filtered one in parallel to demonstrate that any effect observed is due to physical effects and not toxicity) and one control group was tested in the definitive study in a semi-static system.

OBSERVATIONS
The observations of fish were carried out in the following intervals: at 3, 6, 24, 48, 72 and 96 hours. Mortality or any sub-lethal effects were not observed during the experiment.
The test conditions (pH, temperature, oxygen saturation) were measured daily during the test.
The body weights of the fish were recorded before the introduction of fish. The body weight of 7 fish per aquarium was registered. The loading of the aquaria was calculated on the basis of these body weights (1.0 g fish/litre testing liquid).

ANIMAL WELFARE
After the experiment, all fish were humanely killed. Euthanasia for all animals was applied by an overdose of an aqueous solution of 250 mg/L Tricaine Methanesulphonate (MS 222; CAS No.: 886-86-2). This technique is considered to provide a humane euthanasia by direct depression of the brain and vital centres.

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

NOELR

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:

LOELR

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:

LL50

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:

LL100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

VALIDITY
The achievable validity criteria were within acceptable limits and therefore the study can be considered as valid.

MORTALITY DATA
No sublethal effect was observed.

BODY WEIGHT
There was no considerable difference observed concerning body weights between the groups.

Results with reference substance (positive control):

Not required

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects.
The LL50, NOELR, LOELR and LL100 were determined directly from the raw data.

Sublethal observations / clinical signs:

Results of the Preliminary Range-Finding Test

Nominal concentrations [mg/L nominal loading rate WAF]	Untreated control	0.1	1.0	10.0	100.0	100.0*
Number of treated animals	4	4	4	4	4	4
Number of dead animals	0	0	0	0	0	0

*Remark: filtered test solution

 

Cumulative mortality data in the Limit Test

Test group	Cumulative Mortality (number of dead fish) [initial population = 7 fish]
	3 h	6 h	24 h	48 h	72 h	96 h
Control	0	0	0	0	0	0
100.0 mg/L nominal loading rate (unfiltered)	0	0	0	0	0	0
100.0 mg/L nominal loading rate (filtered)	0	0	0	0	0	0

 

Measured and calculated data of bodyweight

Test group	Measured weight of 7 fish (g)	Calculated mean weight of 1 fish (g)	Loading of testing aquarium (g fish/L testing liquid)
Control	1.55	0.22	0.52
100.0 mg/L nominal loading rate (unfiltered)	1.76	0.25	0.59
100.0 mg/L nominal loading rate (filtered)	1.64	0.23	0.55

 

DATA OF TEST CONDITIONS

Temperature

Test group	Measuring (°C)
	0h^1)	24h^2)	24h^1)	48h^2)	48h^1)	72h^2)	72h^1)	96h^2)
Control	22.5	21.9	21.6	20.9	22.4	21.3	22.4	21.1
100.0 mg/L nominal loading rate (unfiltered)	23.5	21.4	21.2	21.0	22.8	21.1	23.1	21.1
100.0 mg/L nominal loading rate (filtered)	23.5	21.4	21.2	21.4	22.8	21.0	23.1	21.1

Dissolved oxygen concentration

Test group	Measuring (%)
	0h^1)	24h^2)	24h^1)	48h^2)	48h^1)	72h^2)	72h^1)	96h^2)
Control	100	89	100	92	98	93	100	91
100.0 mg/L nominal loading rate (unfiltered)	100	94	81	85	93	79	94	90
100.0 mg/L nominal loading rate (filtered)	82	95	73	92	77	95	78	93

pH

Test group	Measuring
	0h^1)	24h^2)	24h^1)	48h^2)	48h^1)	72h^2)	72h^1)	96h^2)
Control	7.83	7.94	8.33	8.18	8.22	7.86	8.09	8.14
100.0 mg/L nominal loading rate (unfiltered)	8.07	8.21	8.14	8.27	8.26	8.16	8.20	8.29
100.0 mg/L nominal loading rate (filtered)	8.20	8.32	8.15	8.28	8.30	8.15	8.19	8.30

¹⁾: measurement at the start of the renewal period in the fresh test medium

²⁾: measurement at the end of the renewal period in the old test medium

 

BODY LENGTH OF TEST FISH

Body length of test fish at the end of the test

Test group	Body length [cm]
	1	2	3	4	5	6	7
Control	2.1	2.5	2.3	2.2	2.5	2.6	2.6
100.0 mg/L nominal loading rate (unfiltered)	2.3	2.4	2.2	2.69	2.5	2.2	2.5
100.0 mg/L nominal loading rate (filtered)	2.2	2.4	2.5	2.5	2.6	2.6	2.4

Remark: AT the end of the test body length of test fish was measured in order to check their compliance with the size range recommended for the species by the test guideline [Commission Regulation (EC) No 440/2008; Annex Part C. C.1]

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of SynNova Base Oil was assessed with acute fish toxicity test on Zebrafish (Brachydanio rerio), over an exposure period of 96 hours in a semi-static system.

Under the conditions of this acute fish toxicity study on Zebrafish (Brachydanio rerio) the observed and calculated endpoints for the effect of SynNova Base Oil were the following:

Based on the nominal loading rate (WAF):
The 24h, 48h, 72h and 96h LL50 value: > 100.0 mg/L
The 96h LL100 value: > 100.0 mg/L
The 96h No-Observed Effect Loading Rate (NOELR): 100.0 mg/L
The 96h Lowest Observed Effect Loading Rate (LOELR): > 100.0 mg/L

Executive summary:

The acute toxicity of SynNova Base Oil was assessed with acute fish toxicity test on Zebrafish (Brachydanio rerio), over an exposure period of 96 hours in a semi-static system.

 

Because no toxic response was observed during the preliminary range-finding test, only a Limit test was carried out using only one nominal test concentration at 100.0 mg/L nominal loading rate WAF (a filtered and a non-filtered one in parallel) and one control group in the definitive study under semi-static conditions.

 

The WAF solution was prepared with direct addition of the test item. During the preparation periods, droplets in the top layers were observed in the test solution, so the solution in the middle layer was collected and siphoned into another test vessel as exposure solution, certainly, there no obvious droplets were observed in the exposure solution. Any remaining non-dissolved test chemical component in the WAF has the possibility to cause physical effects on the test organism unrelated to inherent chemical toxicity. For this reason, a filtered and a non-filtered test item group was tested in parallel to demonstrate that any effect observed is due to physical effects and not toxicity.

 

The test concentration was analytically determined at the start and at the end of the first and the last renewal periods. The measured concentration was below the Limit of Detection (LOD = 0.096 mg/L) at the non-filtered sample, while it was below the Limit of Quantification (LOQ = 0.28 mg/L) at the filtered test item group during the first renewal period. The measured concentration was below the Limit of Detection in both the filtered and non-filtered test item groups during the last renewal period. The biological results are based on the nominal loading rate (WAF).

 

One aquarium was used in the test groups and one in the control group. Each aquarium comprised seven fish and three litre test solution.

 

All achievable validity criteria were met during this study.

 

Under the conditions of this acute fish toxicity study on Zebrafish (Brachydanio rerio) the observed and calculated endpoints for the effect of SynNova Base Oil were the following:

 

Based on the nominal loading rate (WAF):

The 24h, 48h, 72h and 96h LL50 value: > 100.0 mg/L

The 96h LL100 value: > 100.0 mg/L

The 96h No-Observed Effect Loading Rate (NOELR): 100.0 mg/L

The 96h Lowest Observed Effect Loading Rate (LOELR): > 100.0 mg/L

Description of key information

Based on the nominal loading rate (WAF):

The 24h, 48h, 72h and 96h LL50 value: > 100.0 mg/L

The 96h LL100 value: > 100.0 mg/L

The 96h No-Observed Effect Loading Rate (NOELR): 100.0 mg/L

The 96h Lowest Observed Effect Loading Rate (LOELR): > 100.0 mg/L

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

100 mg/L

Additional information

The acute toxicity of SynNova Base Oil was assessed with acute fish toxicity test on Zebrafish (Brachydanio rerio), over an exposure period of 96 hours in a semi-static system.

The WAF solution was prepared with direct addition of the test item. During the preparation periods, droplets in the top layers were observed in the test solution, so the solution in the middle layer was collected and siphoned into another test vessel as exposure solution, certainly, there no obvious droplets were observed in the exposure solution. Any remaining non-dissolved test chemical component in the WAF has the possibility to cause physical effects on the test organism unrelated to inherent chemical toxicity. For this reason, a filtered and a non-filtered test item group was tested in parallel to demonstrate that any effect observed is due to physical effects and not toxicity.

 

Under the conditions of this acute fish toxicity study on Zebrafish (Brachydanio rerio) the observed and calculated endpoints for the effect of SynNova Base Oil were the following:

Based on the nominal loading rate (WAF):

The 24h, 48h, 72h and 96h LL50 value: > 100.0 mg/L

The 96h LL100 value: > 100.0 mg/L

The 96h No-Observed Effect Loading Rate (NOELR): 100.0 mg/L

The 96h Lowest Observed Effect Loading Rate (LOELR): > 100.0 mg/L