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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Sept. 6, 1996-Dec. 9, 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study.
Justification for type of information:
Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, aromatics (2-25%) straddles Hydrocarbons, C9-C14, Aliphatics, (2-25% Aromatics) and Hydrocarbons, C14-C20, Aliphatics (2-30% Aromatics). Read across justification documents have been provided for the same in Section 13 of the dossier. For this substance, a worst case scenario approach has been used for each endpoint.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
Two dissolved oxygen readings were outside the guideline limit. During acclimation, 49 hourly temperature readings were marginally lower than the preferred range of 13-17 degree C. These deviations had no effects on results.
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
- Concentrations: all concentrations
- Sample storage conditions before analysis: If samples could not be analysed the day they were taken, they were stored at 4°C and extracted at the earliest opportunity. Samples were stable for up to 7 days.
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Test substance was added to measured volumes of test medium. The vessels were then sealed, leaving a small headspace, and the contents stirred with a 1-2 cm vortex depth until the aqueous and test substance phases equilibrated (21-22 hrs). The vessels were then allowed to settle for 1.5-2.5 hrs and the aqueous phase drawn off for use in the test as a Water Accomodated Fraction (WAF). The control medium was treated in the same manner.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Strain: Walbaum
- Source: Brow Well Fisheries, Hebden, Yorkshire
- Length at study initiation (length definition, mean, range and SD): 4.3-4.4 cm
- Weight at study initiation (mean and range, SD): 0.68-0.85 g

ACCLIMATION
- Acclimation period: 9 days
- Acclimation conditions (same as test or not): minimum of 7 days at test conditions
- Type and amount of food: Mainstream Trout Food No. 03 (BP Nutrition Ltd.)
- Feeding frequency: maintainence ration
- Health during acclimation (any mortality observed): If mortality is more than 5% in the 7 days preceeding a study, the fish are not used.
Test type:
semi-static
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
none
Hardness:
146-170 mg/l CaCO3
Test temperature:
14.9-16.4°C
pH:
7.4-7.6
Dissolved oxygen:
6.1-9.6
Nominal and measured concentrations:
Nominal loading rate: 0, 0.3, 1, 3, 10, 30, 100 mg/l
Measured: <0.05, 0.05, 0.16, 0.33, 1.2, 2.9. 5.1 (single measure) mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: sealed with a glass plate
- Material, size, headspace, fill volume: 1 l glass bottles completely filled with test substance
- Renewal rate of test solution (frequency/flow rate): fresh test media was added after every observation
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: 500 ft deep Stanlow Well Number 7 treated by softening ion exchange followed by filtration through a twin bed active carbon filter. Water is further filtered after entering the laboratory.
- Intervals of water quality measurement: six months


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations for mortality and toxic signs were made at 3, 24, 48, 72, and 96 hrs.
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
10 - 30 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Key result
Duration:
96 h
Dose descriptor:
NOELR
Effect conc.:
0.3 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
24 h
Dose descriptor:
LL50
Effect conc.:
30 - 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LL50
Effect conc.:
10 - 30 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LL50
Effect conc.:
10 - 30 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities: Yes, some fish at the 10 mg/l concentration showed symptoms of toxicity at the 24 hr observation such as increased cough frequency, hyperventalism, color changes, and swimming abnormally which continued throughout the test. Some fish in the 30 mg/l group displayed abnormal swimming at the 3 and 24 hr observations. At the 48 hr observation all fish in this group were dead. All fish in the 100 mg/l group showed the above symptoms at the 3 hr observation. By the 24 hr observation, all fish in this group were dead except for 1 fish which was dead at the 48 hr observation.
Sublethal observations / clinical signs:

Mortality of O. mykiss

Loading Rate (mg/l)

3 hrs

24 hrs

48 hrs

72 hrs

96 hrs

0

0

0

0

0

0

0.3

0

0

0

0

0

1

0

0

1

1

3

3

0

1

3

3

3

10

0

1

1

1

1

30

0

2

7

7

7

100

0

6

7

7

7

Validity criteria fulfilled:
yes
Conclusions:
The 96-hr LL50 range for Oncorhynchus mykiss was 10-30 mg/l (WAF).
Executive summary:

The purpose of this study was to determine the acute toxicity of white spirits to the fish Oncorhynchus mykiss. Groups of 7 fish were exposed to WAF loading rates of 0, 0.3, 1, 3, 10, 30, and 100 mg/l of test substance for 96 -hrs. The test solutions were renewed daily. Fish were observed at 3, 24, 48, 72, and 96 hrs after start of exposure for mortality and toxic effects. There was some mortality of fish in the 1, 3, and 10 mg/l groups. There was mortality of all animals in the 30 -100 mg/l groups. Behavioural abnormalties were seen at concentrations of 10 mg/l and above. The 96-hr LL50 range for Oncorhynchus mykiss was 10-30 mg/l (WAF). 

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
from 1994-04-07 to 1994-07-15
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study.
Justification for type of information:
Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, aromatics (2-25%) straddles Hydrocarbons, C9-C14, Aliphatics, (2-25% Aromatics) and Hydrocarbons, C14-C20, Aliphatics (2-30% Aromatics). Read across justification documents have been provided for the same in Section 13 of the dossier. For this substance, a worst case scenario approach has been used for each endpoint.
Reason / purpose for cross-reference:
read-across: supporting information
Reason / purpose for cross-reference:
reference to same study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
Water hardness higher than recommended; However, it is assumed that it had no influence on the test results.
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
Samples were taken during the equilibration phase (WAF preparation: media allowed to settle for about 1 hour) and at the end of the period of use 24 hours later.
All extractions for each test were performed on the day of the samples being taken and all determinations carried out on the same or following day.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Water accomodated fractions of Shellsol H were produced by stirring water appropriate to the test organisms and Shellsol H from 69-71 hours, leaving the contents of the vessel to stand at least for 1 hour and running off the aqueous phase for use as the test medium.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Source: Exmoor trout farm, Molton, Devon UK
- Length at study initiation (length definition, mean, range and SD): 4.7-5.2 cm (mean length: 4.9 cm)
- Weight at study initiation (mean and range, SD): 0.87-1.2 g (mean weight: 1.0 g)

ACCLIMATION
- Acclimation period: at least 9 days
- Acclimation conditions (same as test or not): minimum of 7 days at test conditions
- Type and amount of food: Mainstream Trout Food No. 03 (BP Nutrition Ltd.)
- Feeding frequency: maintenance ration
- Health during acclimation (any mortality observed): If mortality is more than 5% in the 7 days preceeding a study, the fish are not used.
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
none
Hardness:
258-254 mg/l CaCO3
Test temperature:
14-15°C
pH:
7.1-7.2
Dissolved oxygen:
9.4-10.1
Salinity:
Not applicable (freshwater)
Nominal and measured concentrations:
Nominal loading rate: 0, 10, 100, 1000 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: sealed with a glass plate
- Material, size, headspace, fill volume: 11 l glass aspirators completely filled with test medium (WAF) or dilution watr (control)
- Renewal rate of test solution (frequency/flow rate): fresh test media was added after every observation
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water comes from two pumping stations (Highstead and controlled by the Mid Kent Water Company. The water is obtained from bore holes in the chalk of the North Downs. The only chemical treatment prior to its arrival in the laboratory is chlorination to 0.1 mg/l. In the laboratory, the water is filtered (PAL MDY 1001 Y400) to remove all particles larger than 15 µm (90% of particles greater than 10µm) and passed through activated carbon filters (Cuno model CT) to remove chlorine and organic conatminants. Both particle and activated carbon filter cartridges are renewed as recommended by the manufacturers. Heat exchange units (stainless steel and perspex) are used to adjust the temperature of the water.
- Intervals of water quality measurement: six months


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations for mortality and toxic signs were made at 24, 48, 72, and 96 hrs.
Reference substance (positive control):
not specified
Key result
Duration:
96 h
Dose descriptor:
LL50
Effect conc.:
10 - 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
24 h
Dose descriptor:
LL50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
48 h
Dose descriptor:
LL50
Effect conc.:
100 - 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
72 h
Dose descriptor:
LL50
Effect conc.:
10 - 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Validity criteria fulfilled:
yes
Conclusions:
The 96-hr LL50 range for Oncorhynchus mykiss was 10-100 mg/l (WAF).
Executive summary:

The acute toxicity of water accomodated fractions (aqueous extrats) of Shellsol H has been determined to the fish Oncorhynchus mykiss (rainbow trout).

The test media were prepared by adding quantities of Shellsol H to culture medium appropriate to the test organisms and stirring in sealed vessels for 69 -71 hours). The strirring period was demonstrated as being sufficient to ensure equilibration of the shellsol H and aqueous phases. After stirring the contents of the test vessels were left to settle for about 1 hour before drawing off the WAF for use as the test media in the toxicity tests.

The concentrations of dissolved hydrocarbons (derived from the Shellsol H) in the test media was determined during the toxicit tests usig gas chromatography. The analyses showed that the concentration in the test media was <1% of the loading rate. The reduction in the concentration of dissolved hydrocarbons in the test media during the test was ≤26%.

The acute toxicity to O. mykiss of water accomodated fractions of Shellsol H prepared at loading rates of 10, 100, and 1000 mg/l was determined in a 96h semi-static toxicity test with daily renewal of the test media. The test was conducted in sealed test vessels. The 24, 48, 72 and 96 h LL50 values (loading rates resulting in mortality of 50% of the fish after 24, 48, 72 and 96 h exposure) were >1000, 100 -1000, 10 -100 and 10 -100 mg/l respectively.

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
The study was given a reliability rating of 1 because it followed a standard test guideline, which describes a procedure to evaluate this endpoint, and the results were reviewed for reliability and assessed as valid. The study was also conducted under GLP.
Justification for type of information:
Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, aromatics (2-25%) straddles Hydrocarbons, C9-C14, Aliphatics, (2-25% Aromatics) and Hydrocarbons, C14-C20, Aliphatics (2-30% Aromatics). Read across justification documents have been provided for the same in Section 13 of the dossier. For this substance, a worst case scenario approach has been used for each endpoint.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
An equilibration trial was performed to determine the time required for the components of the test substance to achieve maximum aqueous solubility. Five glass aspirators were each filled with 22L of main dechlorinated water, leaving a small headspace of air. The test substance was added to four of the vessels at a loading rate of 1000 mg/L, and the contents of the fifth aspirator served as the control. All five vessels were then sealed, and stirred as vigorously as possible without creating a dispersion in the sealed vessels, which contained a small headspace of air to permit stirring. After nominal stirring periods of 24, 48, 72, 96and 198 hours, the WAF solutions were allowed to settle for 1-3 hours, after which a sample of the aqueous layer was extracted and analyzed by GC/FID. The results of the equilibration study showed that equilibrium between the test substance components and the aqueous phase at a loading of 1 g/L was reached in about 72-96 hours.
Vehicle:
no
Details on test solutions:
For purposes of the fish toxicity test, individual treatment solutions were prepared as Water Accommodated Fractions (WAFs). This included adding test substance to 15-16 liter glass aspirator bottles, which contained laboratory dilution water, that was filtered (15 um) through
activated charcoal. The WAF was stirred for 93-96 hours as vigorously as possible without creating a dispersion in the sealed vessels, which contained a small headspace of air to permit stirring. After mixing, the WAFs were allowed to settle for one hour, after which the aqueous phase was removed for testing.

A limit test was conducted using one replicate each for a control WAF, and at a 1000 mg/L WAF loading concentration. The loading rate was based on the results of a range-finding study, details of which were not included in the definitive study report. Each replicate system contained seven fish in completely filled (11L of WAF or control water) glass aspirator bottles that were sealed with glass plates to exclude air. After 24, 48 and 72 hours exposure any dead or immobilized fish were removed and surviving fish were transferred to previously prepared WAFs. After 96 hrs exposure, the fish were observed again and the test was terminated. Samples of each batch of WAF were taken at the time of preparation and at the end of the period of use 24 hours later for GC/FID analysis, D.O. and pH. Temperature was measured hourly in a vessel adjacent to the test vessels throughout the test.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
Fish were obtained from Hartford Trout Farm, Brompton Regis, Somerset U.K. and acclimated to the test conditions for a minimum of seven days before use. A sample of seven of the fish used in the test had a mean length of 4.6 cm and a mean weight of 0.96 g. The fish were not fed in the 24 hr preceding the test, nor during the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Hardness:
Water hardness was 244 to 268mg/L as CaCO3.
Test temperature:
Test temperature was 15.3-15.9 Deg C.
pH:
The pH ranged from 7.2 to 7.5 during the study.
Dissolved oxygen:
Dissolved oxygen was 8.9 to 9.8 mg/L.
Salinity:
freshwater
Nominal and measured concentrations:
WAF concentration (mg/L) of dissolved hydrocarbon (derived from test substance) as determined by GC/FID. Specific hydrocarbon isomers were not identified. Analytical results are based on mass attributable to the test material. Control concentration was <0.05 mg/L at all sampling intervals identified above.

Time 1000 mg/L WAF
0 hr 0.31
24 hr new 0.35
24 hr old 0.17
48 hr new 0.28
48 hr old 0.19
72 hr new 0.35
72 hr old 0.20
96 hr old 0.26
Mean 0.26
Details on test conditions:
Artificial light cycle was 16 hours on and 8 hours dark.
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LL0
Effect conc.:
>= 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
No mortality observed during the 96 hour test.

96-hour LL0 = 1000 mg/L
96 hour LL50 >1000 mg/L

An LC50 based upon measured values was not calculated.

This value is representative of acute aquatic fish toxicity for those C14-C20 (high aromatics) aliphatic category members having an initial minimum boiling point range of 264C. Since the hydrocarbon constituents for these category members with boiling point values greater than 264 deg C do not show adequate water solubility in equilibrated aqueous solutions, acute aquatic toxicity effects are not expected at the 1000 mg/L maximum loading concentration investigated in this study
Reported statistics and error estimates:
No statistics performed.
Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%), as measured by mortality to rainbow trout (Oncorhynchus mykiss) was evaluated in freshwater. Under the conditions of this study, Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%) did not produce acute lethal toxicity to Oncorhynchus mykiss at 1000 mg/l, based on nominal loading of the test substance in water.
Executive summary:

The acute toxicity of Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%), as measured by mortality to rainbow trout (Oncorhynchus mykiss) was evaluated in freshwater. Under the conditions of this study, Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%) did not produce acute lethal toxicity to Oncorhynchus mykiss at 1000 mg/l, based on nominal loading of the test substance in water.

Description of key information

There is no experimental data available for Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, 2-25% aromatics. Data is read across from structural analogues such as Hydrocarbons, C9-C14 aliphatics, 2-25% aromatics and Hydrocarbons, C14-C20 aliphatic, 2-30% aromatics.

Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, aromatics (2-25%) straddles Hydrocarbons, C9-C14, Aliphatics, (2-25% Aromatics) and Hydrocarbons, C14-C20, Aliphatics (2-30% Aromatics). Read across justification documents have been provided for the same in Section 13 of the dossier. For this substance, a worst case scenario approach has been used for each endpoint.

The acute toxicity of Hydrocarbons, C9-C12, n-alkanes, isoalkanes, cyclics, aromatics (2-25%) as measured by mortality to rainbow trout (Oncorhynchus mykiss) was evaluated in freshwater. Groups of 7 fish were exposed to WAF loading rates of 0, 0.3, 1, 3, 10, 30, and 100 mg/L of test substance for 96-hrs. There was some mortality of fish in the 1, 3, and 10 mg/L groups. There was mortality of all animals in the 30 -100 mg/L groups. Behavioural abnormalities were seen at concentrations of 10 mg/L and above. The 96-hr LL50 range for Oncorhynchus mykiss was 10-30 mg/L (WAF). 

 

The acute toxicity of Hydrocarbons, C10-C13, n-alkanes, isoalkanes, cyclics, aromatics (2-25%) as measured by mortality to rainbow trout (Oncorhyncus mykiss) was evaluated in freshwater. The 96-hr LL50 range for Oncorhynchus mykiss was 10-100 mg/L (WAF). 

 

The acute toxicity of Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%), as measured by mortality to rainbow trout (Oncorhychus mykiss) was evaluated in freshwater. Under the conditions of this study, Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%) did not produce acute lethal toxicity to Oncorhynchus mykiss at 1000 mg/L, based on nominal loading of the test substance in water.

 

In another study, the acute toxicity of Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%), as measured by mortality to rainbow trout (Oncorhynchus mykiss) was evaluated in freshwater. Under the conditions of this study, Hydrocarbons, C16-C20, n-alkanes, isoalkanes, cyclics, aromatics (2-30%) did not produce acute lethal toxicity to Oncorhynchus mykiss at 1000 mg/L, based on nominal loading of the test substance in water.

These data are used as read across data for Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, 2-25% aromatics.

Key value for chemical safety assessment

Additional information

The estimated freshwater fish 96 hr LL50 value for Hydrocarbons, C12-C16, n-alkanes,isoalkanes, cyclics, aromatics (2-25%) determined using the Petrotox computer model (CONCAWE, 2010) is 1117.88 mg/L with rainbow trout, Oncorhynchus mykiss based on mortality.

Water accommodated fractions of Hydrocarbons, C9-C12, n-alkanes, isoalkanes, cyclics, aromatics (2 -25%), produced a 96-hour LL50 value in the range 10-30 mg/L with rainbow trout, Oncorhynchus mykiss.

 

Water accommodated fractions of Hydrocarbons, C10-C13, n-alkanes, isoalkanes, cyclics, aromatics (2 -25%), produced a 96-hour LL50 value in the range 10-100 mg/L with rainbow trout, Oncorhynchus mykiss.

 

Water accommodated fractions of hydrocarbons, C16 -C20, n-alkanes, isoalkanes, cyclics, aromatics (2 -30%) did not cause acute toxicity to fish at a nominal loading of 1000 mg/L with rainbow trout, Oncorhynchus mykiss.

 

These data are used as read across data for Hydrocarbons, C12-C16, n-alkanes, isoalkanes, cyclics, 2-25% aromatics.