Tar bases, coal, lutidine fraction

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:

Experimental start: 04 March 2016, Experimental termination: 01 April 2016, Finalisation: 10 May 2016

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:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: WC 2015.10533
- Expiration date of the lot/batch: 31 August 2017
- Purity test date: 99.3% w/w

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: room temperature in the dark
- Stability under test conditions: yes
- Solubility and stability of the test substance in the solvent/vehicle: yes
- Reactivity of the test substance with the solvent/vehicle: no

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: no
- Preliminary purification step (if any): no

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The concentrations of the test substance were analysed in each treatment group.
- Sampling method: Duplicate samples were taken from each test vessel at the start of the test and after 24 hours from the 100 mg/L test concentration and after 96 hours from the 6.25, 12.5, 25 and 50 mg/L test concentrations.
- Sample storage conditions before analysis: all samples were stored frozen immediately after sampling.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
A concentrated stock solution of nominal 1000 mg/L was freshly prepared by mixing 5 g of the test item into 5 litres of test water by vigorously stirring for 15 minutes with a magnetic stirrer. Adequate volumes of the stock solution were added to test water in the test vessels and were mixed for 2 minutes with a flat-bladed stirrer to prepare the test media with the test concentrations of 6.25, 12.5, 25, 50 and 100 mg/L.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: Oncorhynchus mykiss
- Source: Brow Well Fisheries Limited, Hebden, Yorkshire, United Kingdom
- Age at study initiation (mean and range, SD): not given
- Length at study initiation (length definition, mean, range and SD): not given
- Weight at study initiation (mean and range, SD): not given

ACCLIMATION
- Acclimation period: one week
- Acclimation conditions (same as test or not): under test conditions
- Type and amount of food during acclimation: commercial trout pellets
- Feeding frequency during acclimation: not given
- Health during acclimation (any mortality observed): all fish were healthy and mortality was less than 5%

FEEDING DURING TEST
- Not applicable, feeding was discontinued approximately 22.5 hours prior to the start of the definitive test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Hardness:

140 mg/L as CaC03

Test temperature:

14 °C

pH:

7.2 to 8

Dissolved oxygen:

5.4 to 11.2 mg/L

Salinity:

not reported

Conductivity:

392.34 µS/cm at 20 °C

Nominal and measured concentrations:

Nominal concentrations: 0, 6.25, 12.5, 25, 50 and 100 mg/L
Measured concentrations: measured concentrations ranged from 88 to 99% of nominal concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel: a glass vessel (fish bowl with ca 33 cm diameter) with 20 litres of test medium was used for each treatment
- Type (delete if not applicable): open
- Material, size, headspace, fill volume:
- Aeration: during the test, the test media was left unaerated, except after observations at 48 and 72 hours when 45 and 30 minutes, respectively, of aeration was provided to increase the dissolved oxygen concentrations of the test solutions
- No. of organisms per vessel: 7 fish were introduced in each test vessel at random order
- No. of vessels per concentration (replicates): one test vessel per concentration and control
- No. of vessels per control (replicates): one control was tested in parallel
- No. of vessels per vehicle control (replicates):
- Biomass loading rate: 0.33 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water was dechlorinated by passage through activated carbon filter and partly softened, then passed through a series of computer controlled plate heat exchangers to achieve the required temperature
- Total organic carbon: 1.06 mg/L
- Particulate matter: <0.089 NTU
- Metals: not given
- Pesticides: 0.12 µg/L
- Chlorine: 0.245 mg/L free, 0.33 mg/L total
- Alkalinity: not given
- Ca/mg ratio: not given
- Culture medium different from test medium: no
- Intervals of water quality measurement: not given

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hour light, 8 hour darkness cycle
- Light intensity: in the range from 259 to 332 Lux

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline:
- Range finding study: A limit test was conducted at 100 mg/L, as the EC50 values obtained in the algal and daphnia study were >100 mg/L. However, 100% mortality was observed after 3 hours.
- Test concentrations: Subsequently, a range finding test with concentrations of 1, 10 and 100 mg/L was carried out.
- Results used to determine the conditions for the definitive study: No mortality was seen at 1 and 10 mg/L, whereas 100% mortality occurred at 100 mg/L.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

71 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No mortality occurred up to the test concentration of 50 mg/L, whereas 100% mortality occurred at the test concentration of 100 mg/L.

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

Not applicable

Sublethal observations / clinical signs:

Table 1: Mortality and visible abnormalities observed in the test fish

Nominal test concentration (mg/L)	Number of fish showing abnormalities/Cumulative dead fish
	Observation time
	1 hour	3 hours	6 hours	24 hours	48 hours	72 hours	96 hours
Control	0/0	0/0	0/0	0/0	0/0	0/0	0/0
6.25	0/0	0/0	0/0	0/0	0/0	0/0	0/0
12.5	0/0	0/0	0/0	0/0	0/0	0/0	0/0
25	0/0	0/0	0/0	0/0	0/0	0/0	0/0
50	0/0	0/0	0/0	0/0	0/0	0/0	0/0
100	7/0	7/0	7/7	-	-	-	-

Validity criteria fulfilled:

yes

Conclusions:

This static acute aquatic toxicity study with rainbow trout resulted in a 96-hour LC50 value of 71 mg/L.

Executive summary:

The acute toxicity of the test substance to rainbow trout (Onchorhynchus mykiss) was studied in a static test over 96 hours in according to OECD TG 203 under GLP. The experiment is considered relevant, adequate and conclusive.

Seven fish per dose group (loading rate of 0.33 g/L) were exposed in a glass vessel of ca. 33 cm diamter in a volume of 20 L test medium. Based on a range finding study, nominal concentrations of 0 (control), 6.25, 12.5, 25, 50 and 100 mg/L were tested, and test solutions were prepared from a freshly prepared stock solution. Analysis of the test concentrations by HPLC with UV detection showed that the measured concentrations were in the range of 88 to 99% of nominal concentrations, and effect levels thus were based on nominal concentrations. No mortalities or other abnormal effects occurred over the test period of 96 hours up to the test concentration of 50 mg/L, whereas 100% mortality was observed at the highest test concentration of 100 mg/L after 6 hours.

Description of key information

96-hour LC50 = 71.0 mg/L, rainbow trout, Sacker 2016 (OECD TG 203, GLP)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

71 mg/L