Non-graphitic carbon fibre

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:

2012-10-01 to 2013-02-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

carbon

EC Number:

701-026-1

Cas Number:

7440-44-0

Molecular formula:

C up to (C80N5H16O)n

IUPAC Name:

carbon

Test material form:

solid: fibres

Details on test material:

- milled carbonised PAN based fibre (Sigrafil C30 M150 UNS)
- for further details on the test material, see section 1.4, endpoint "Non-graphitic carbon fibre (carbonised PAN based fibre, milled)", attached document "Analytics_milled_carbonised_PAN_based_fibre.pdf"

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Water accomodated fraction (WAF) according to OECD guideline 23
- Eluate: not applicable
- Differential loading: no
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): yes

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish (Danio rerio)
- Strain: not reported
- Source: Test facility bred
- Age at study initiation (mean and range, SD): not reported
- Length at study initiation (length definition, mean, range and SD): 2.0 ± 1 cm
- Weight at study initiation (mean and range, SD): 0.11 ± 0.02 g; range: 0.08-0.14 g

- Method of breeding: not reported
- Feeding during test: no
- Food type: live brine shrimp (Artemia spp.) nauplii and ground flake food TetraMinR (Tetra Werke, Melle, Germany)
- Amount: ad libitum
- Frequency: once per day

ACCLIMATION
- Acclimation period: at least 12 days prior to test
- Acclimation conditions (same as test or not): same
- Type and amount of food: ad libitum: live brine shrimp (Artemia spp.) nauplii and ground flake food TetraMinR (Tetra Werke, Melle, Germany)
- Feeding frequency: once per day
- Health during acclimation (any mortality observed): less than 5% in the week preceding the start of the study

QUARANTINE (wild caught): not applicable

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

not reported

Test conditions

Hardness:

total hardness: 1.1 mmol/L

Test temperature:

23 ± 2 °C

pH:

8.5 - 8.6

Dissolved oxygen:

96 - 100%

Conductivity:

238.1 µS/cm

Nominal and measured concentrations:

nominal concentration: 100 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: full glass aquaria
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: glass, 10 L
- Aeration: slightly aerated via a glass capillary
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): no
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 7
- No. of vessels per vehicle control (replicates): not applicable
- Biomass loading rate: not reported

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified tap water, filtration through activated charcoal, passage through lime stone, and aeration
- Total organic carbon: not reported
- Particulate matter: not reported
- Metals: reported
- Pesticides: not reported
- Chlorine: <0.02 mg/L
- Alkalinity: 2.0 mmol/L
- Ca/Mg ratio: 0.8 mmol/L / 0.3 mmol/L
- Conductivity: 238.1 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: not reported

OTHER TEST CONDITIONS
- Adjustment of pH: not reported
- Photoperiod: 12/12


TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: No mortality, clinical signs or any distress were observed throughout the range finding test
- Range finding study: performed prior to main test
- Test concentrations applied in range finding study: 1 mg/L, 10 mg/L, 100 mg/L
- Test concentration applied in the main study: 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

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: no clinical signs and no mortality observed

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LL0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: not detected
- Observations on body length and weight: not reported
- Other biological observations: not detected
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: not reported
- Effect concentrations exceeding solubility of substance in test medium: yes

Results with reference substance (positive control):

No reference substance tested

Any other information on results incl. tables

Sublethal observations / clinical signs:

Cumulative mortality and clinical signs during the test period of 96 h (number of fish per treatment: 7)

Nominal concentration of the carbon fibre [mg/L]	Test duration
	3 h	24 h	48 h	72 h	96 h
Control	nd.	nd.	nd.	nd.	nd.
100	nd.	nd.	nd.	nd.	nd.

nd. = no symptoms detected

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the 96 h acute toxicity test, conducted according to OECD guideline 203 and EU method C.1, zebrafish (Danio rerio) were exposed to the test substance (milled carbonised PAN based fibre) at a nominal loading concentration of 100 mg/L under static conditions for the period of 96 hours. Neither clinical signs nor mortality could be observed throughout the study.
Based on the results of this study, the LL50 of the test substance was determined to be > 100 mg test item/L. The NOEC based on the visual observation is reported as >= 100 mg/L (NOEL >= 100 mg/L).
This toxicity study is classified as acceptable and satisfies the guideline requirement for “Fish, Acute Toxicity Test”.

Executive summary:

A study was performed at the Fraunhofer Institute for Molecular Biology and Applied Ecology to determine the acute toxicity of the test item (milled carbonised PAN based fibre) to zebrafish (Danio rerio). The study was conducted according to the OECD guideline 203 and EU method C.1.

Due to the very low water solubility of the test item a Water Accommodated Fraction (WAF) according to OECD guideline 23 was applied. The test item was examined in a limit test at one nominal concentration of 100 mg/L under static conditions for 96 hours. An untreated dilution water control was run in parallel. Seven fish were used at the test concentration and in the control.

There were no toxic effects of the test item on zebrafish (Danio rerio) at the loading of 100 mg test item/L (nominal). Neither clinical signs nor mortality were observed throughout the study. Thus, no toxicity effect value LC₅₀ for acute toxicity was calculated. The 50 % lethal loading (LL50) based on the nominal loading was determined to be > 100 mg/L. The NOEC based on the visual observation of clinical signs and mortality is reported as ≥ 100 mg/L (NOEL ≥ 100 mg/L).