Hydrocarbons, C13-16

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-03-14 to 2002-03-19

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well documented guideline study. The substance was identified by a statement from the manufacturer. No analytical monitoring.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: "Guideline for measuring the acute toxicity of offshore chemicals to juvenile marine fish, the turbot Scophthalmus maximus and the shepshead minnow Cyprinodon variegatus", based on OECD guideline for Testing of Chemicals n°203 (17/07/92)

Deviations:

no

Principles of method if other than guideline:

guideline study

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Hydrocarbons, C14-C18, n-alkanes, isoalkanes, cyclics, <2% aromatics

EC Number:

927-632-8

IUPAC Name:

Hydrocarbons, C14-C18, n-alkanes, isoalkanes, cyclics, <2% aromatics

Constituent 2

Reference substance name:

DEV 2001/37

IUPAC Name:

DEV 2001/37

Details on test material:

- Name of test material (as cited in study report): DEV 2001/37
- Substance type: Petroleum product, UVCB
- Physical state: Colourless liquid
- Analytical purity: 100% Commercial product
- Stability under test conditions: stable

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
No test surrogate material is tested

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

Not applicable (no analytical monitoring)

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accomodation fractions
WAF of different loadings were prepared in 10 L glass flasks by mixing different amounts of test substance with dilution water (natural seawater with a salinityof 32-34 per thousand filtered on a gravel filter to 50-70µm) for 20 hrs. The mixture was sedimented for minimum 2.5 hrs before the WAF were siphoned into the individual test vessels.
- Controls: Dilution water without test substance

Test organisms

Test organisms (species):

other: Scophthalmus maximus

Details on test organisms:

TEST ORGANISM
- Common name: Turbot
- Source: Stolt Sea Farms, Oye, Norway
- Age at study initiation: Juvenile
- Length at study initiation: 40-60 mm.
- Feeding during test: no feeding

ACCLIMATION
- Acclimation conditions: continuous flow of natural gravel filtered sea water at 10 +/- 3°C and salinity of 32-34 per thousand under normal light intensity (40 W tungsten lamps) at 16:8h L:D cycle.
- Type of food: commercila fish fry feed
- Feeding frequency: every 3 days

Study design

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No post exposure observation

Test conditions

Hardness:

Not relevant (seawater study)

Test temperature:

15 +/- 1.5 °C

pH:

7.80 - 8.03

Dissolved oxygen:

85 - 98% saturation

Salinity:

45.0 - 45.5 mS

Nominal and measured concentrations:

Nominal concentrations (based upon loading rate): 27, 66, 164, 410, 1028 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: open
- Material, size, headspace, fill volume: White polyethylene vessels, 35 L, filled with 10 L WAF (test) or dilution seawater (control)
- Aeration: yes
- Renewal rate of test solution: every 48 hrs
- No. of organisms per vessel (test): 7
- No. of organisms per vessel (control): 10
- No. of organisms per vessel (sensitivity control): 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 1
- No. of vessels per sensitivity control (replicates): 1 per concentration
- Biomass loading rate: test 2.91 mg/L; control 1.38 mg/L


TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: Natural seawater with a salinity of 32-34 per thousand filtered in a gravel filter to 50-70 µm.
- Intervals of water quality measurement: every 48 hrs


OTHER TEST CONDITIONS
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 40 W tungsten lamps


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : immobilisation


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 1 028 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0%
- Other adverse effects control: None

Results with reference substance (positive control):

- Results with reference substance valid
- mortality: 20% after 96hr exposure to 0.53 mg/l 3,5-dichlorophenol; 100% mortality after 24hr exposure to 0.67 mg/l 3,5-dichlorophenol

Reported statistics and error estimates:

No calculations possible

Any other information on results incl. tables

Sublethal observations / clinical signs:

Test acceptability:

- Temperature in the exposure vessels during 96 hrs of exposure was 15.0 +/- 0.7 °C.

- The fish in the control series showed normal swimming behaviour showing a cumulative immobilization/mortality of 0% after 96 -hr exposure.

- Saturation of oxygen in the control series and the highest concentration of the test did not fall below 85% during the exposure.

- The fish in the sensitivity control series exposed to 0.53 mg/L of 3.5 -dichlorophenol behaved lethagic but with normal swimming behaviour and a cumulative immobilisation/mortality of 20% after 96 -hr exposure. A second sensitivity control series exosed to 0.67 mg/l of 3,5 -dichlorophenol behaved lethargic after 24 hr exposure with a cumulative immobilization/mortality of 100% after 48 hr exposure.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

-sensitivity of the fish exposed to 0.6 mg/l of 3,5-dichorophenol in the range of 20-80% immobile animals; immobilisation in the control series <= 10% after 96-hr exposure; saturation of oxygen greater than 60%; temperature in the range 15+/-1.5°C

Conclusions:

96h-LL50 > 1028 mg/L (based on nominal loading rate)

Executive summary:

An acute toxicity test was conducted on juveniles of the marine fish Scophthalmus maximus (Turbot). The test substance concentrations (nominal loading rates) ranged from 27 to 1028 mg/L. Test solutions were prepared as individual water accomodated fractions (WAF). Control and dilution water were natural seawater of 32 -34 ⁰/₀₀ salinity filtered through a gravel filter to 50 -70 µm. Test substance was mixed in dilution water for 20 hrs and the mixture was allowed to settle for a minimum of 2.5 hr prior to drawing off the aqueous solution for testing. Test vessels were 35 -L white polyethylene vessels, each containing 10 L of test solution. In the test, 5 concentrations were tested in a geometric series: 27, 66, 165, 410, and 1028 mg/L. Two replicates each containing 7 fishes (biomass loading rate: 2.91 g/L) for the testing and 1 replicate containing 10 fishes (biomass loading rate: 1.38 g/L) for the control were used.

Dissolved oxygen was >60% saturation. The pH was 7.80 - 8.03. The temperature was 15.0 +/- 0.7 °C.

Concentration (mg/L)	Day 0 (initiation)				Day 4 (96 hrs)
	Temperature (°C)	Salinity (mS)	% O2	pH	Temperature (°C)	Salinity (mS)	% O2	pH
Control	15.1	45.0	96	8.01	15.7	45.3	87	7.86
27	-	45.0	93	8.01	-	45.4	90	7.82
66	-	45.1	92	8.02	-	45.3	89	7.81
165	-	45.0	91	8.01	-	45.2	90	7.81
410	-	45.0	90	8.03	-	45.4	91	7.82
1028	-	45.0	90	8.04	-	45.5	92	7.83

Test organisms were obtained from a fish farm (Stolt Sea Farms, Oye, Norway). At the test start, the organisms were 40 - 60 mm long juveniles.

No deviations from the protocol were noted.

Analytical monitoring was not performed. Therefore the results are based on the loading rate.

Nominal loading (mg/L)	Number of fish
	Initial	Immobilised/dead
		24h	48 h	72 h	96 h	Cumulative
Control	10	0	0	0	0	0
27	7 x 2	0	0	0	0	0
66	7 x 2	0	0	0	0	0
165	7 x 2	0	0	0	0	0
410	7 x 2	0	0	0	0	0
1028	7 x 2	0	0	0	0	0

96h-LL50>1028 mg/L (based upon nominal loading rate).