Coffee, bean, roasted, ext.

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:

From 04 to 30 November 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: CF02
- Expiration date of the lot/batch: 31.01.2019
- Purity test date: not available

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at 4°C, in the dark
- Stability under test conditions: not available
- Solubility and stability of the test substance in the solvent/vehicle: water solubility 2.85 mg/L
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not applicable

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Due to the low limit of water solubility of the test item Water accommodated Fraction (WAF) were tested. Supersaturated stock suspensions of the test item were prepared separately by suspending test item in test water at concentrations of 100, 50, 25, 12.5 and 6.25 mg test item/L.
- Preliminary purification step (if any): none
- Final dilution of a dissolved solid, stock liquid or gel: not applicable
- Final preparation of a solid: not applicable

OTHER SPECIFICS: none

Analytical monitoring:

yes

Details on sampling:

Test solutions were sampled from below the water surface via a drain that was approximately 5 cm above the vessel bottom.

Vehicle:

yes

Remarks:

Reconstituted water

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout (Oncorhynchus mykiss)
- Strain: not specified
- Source: Forellenzuchtbetrieb Störk, 88348 Bad Saulgau, Germany
- Age at study initiation (mean and range, SD): not specified
- Length at study initiation (length definition, mean, range and SD): 4.24 ± 0.3 cm
- Weight at study initiation (mean and range, SD): 0.81 ± 0.1 g
- Method of breeding: not specified
- Maintenance of the brood fish: not specified

ACCLIMATION
- Acclimation period: at least 12 days
- Acclimation conditions (same as test or not): held in water of the quality to be used in the test for at least seven days immediately before testing
- Type and amount of food during acclimation: not specified
- Feeding frequency during acclimation: three times per week or daily until 24 hours before the test was started
- Health during acclimation (any mortality observed): no mortality observed

FEEDING DURING TEST: no

Test type:

static

Water media type:

other: Reconstituted water (ISO medium)

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

none

Hardness:

2.5 mmol/L

Test temperature:

14.-16.4°C

pH:

7.5-7.9

Nominal and measured concentrations:

6.25, 12.5, 25, 50 and 100 mg/L loading rate Water Accommodated Fraction

Details on test conditions:

TEST SYSTEM
- Test vessel: aquaria
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 12 liter glass aquaria
- Aeration: 98 to 100 % of the air saturation value
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: not applicable

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water (ISO medium)
- Total organic carbon: 10.721 mg C/L at 0h
- Particulate matter: not specified
- Metals: not specified
- Pesticides: not specified
- Chlorine: not specified
- Alkalinity: 0.8 mmol/L
- Ca/mg ratio: 4:1 (based on molarity)
- Culture medium different from test medium: no
- Intervals of water quality measurement: not specified

OTHER TEST CONDITIONS
- Adjustment of pH: not specified
- Photoperiod: 16h light: 8h dark
- Light intensity: 650 – 730 lux

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: none, a limit test was performed at 100 mg/L WAF prior to dose response test
- Test concentrations: 6.25, 12.5, 25, 50 and 100 mg/L loading rate Water Accommodated Fraction
- Results used to determine the conditions for the definitive study: sub-lethal effects and mortality

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: WAF

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

other: NOEL

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

other: WAF

Basis for effect:

other: sub-lethal effects

Key result

Duration:

96 h

Dose descriptor:

other: LOEL

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

other: WAF

Basis for effect:

other: strong ventilation

Details on results:

- Behavioural abnormalities: some fish remained at the water surface, possibly caused by the coloration of the test medium
- Observations on body length and weight: not specified
- Other biological observations: not specified
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: strong ventilation at 100 mg/L loading rate WAF
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: not specified
- Effect concentrations exceeding solubility of substance in test medium: none

Reported statistics and error estimates:

No statistical analysis was performed

Sublethal observations / clinical signs:

See attached additional information on results.

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity test performed on Rainbow trout, and conducted according to OECD test guideline 203 and to GLP, the NOEL and LOEL values were determined at 50 and 100 mg/L loading rate as a Water Accommodated Fraction (WAF) after 96 hours of exposure to Coffee, bean, roasted, ext. The 96-hour LC50 was determined to be > 100 mg/L loading rate WAF.

Executive summary:

The acute toxicity of Coffee, bean, roasted, ext. to rainbow trout (Oncorhynchus mykiss) was determined in dose response test according to the OECD test guideline 203 and to GLP.

Due to the low aqueous solubility and complex nature of the test item, the test medium was prepared as a Water Accommodated Fraction (WAF).

Seven fish were exposed to a WAF of the test item at nominal loading rates of 0, 6.25, 12.5, 25, 50 and 100 mg/L for 96 hours under static conditions. Morality and sub-lethal effects were recorded after 24, 48, 72 and 96 hours.

No mortality was observed in the control and the test item concentration up to 100 mg/L loading rate WAF. At 100 mg/L loading rate WAF, all fish displayed strong ventilation shortly after insertion into the test concentration.

Some fish were observed mainly at the water surface at 100 mg/L loading rate WAF after 48, 72 and 96 hours. At 72 hours 3 fish were observed mainly at the water surface at 50 mg/L loading rate WAF. The position “mainly at the water surface” was possibly caused by the coloration of the test medium and was not judged as sub-lethal effect and not considered for the NOEL / LOEL determination.

During the test the fish were exposed to 21.1 and 10.3 mg Carbon/L of the nominal loadings of 100 and 50 mg test item/L, respectively. The results for nominal loadings of 25, 12.5 and 6.25 mg Carbon/L were below the Limit of Quantification, however, they were clearly below the NOEL.

The NOEL and LOEL after 96 hours of exposure were determined at 50 and 100 mg/L loading rate WAF respectively.

The 96-hour LC50 was determined to be > 100 mg/L loading rate WAF.

Description of key information

NOEL: 50 mg/L loading rate WAF

LOEL: 100 mg/L loading rate WAF

LC50: >100 mg/L loading rate WAF

Additional information

The acute toxicity of Coffee, bean, roasted, ext. to rainbow trout (Oncorhynchus mykiss) was determined in dose response test according to the OECD test guideline 203 and to GLP.

Due to the low aqueous solubility and complex nature of the test item, the test medium was prepared as a Water Accommodated Fraction (WAF).

Seven fish were exposed to WAFs of the test item at nominal loading rates of 0, 6.25, 12.5, 25, 50 and 100 mg/L for 96 hours under static conditions. Morality and sub-lethal effects were recorded after 24, 48, 72 and 96 hours.

No mortality was observed in the control and the test item concentration up to 100 mg/L loading rate WAF. At 100 mg/L loading rate WAF, all fish displayed strong ventilation shortly after insertion into the test concentration.

Some fish were observed mainly at the water surface at 100 mg/L loading rate WAF after 48, 72 and 96 hours. At 72 hours 3 fish were observed mainly at the water surface at 50 mg/L loading rate WAF. The position “mainly at the water surface” was possibly caused by the coloration of the test medium and was not judged as sub-lethal effect and not considered for the NOEL / LOEL determination.

During the test the fish were exposed to 21.1 and 10.3 mg Carbon/L of the nominal loadings of 100 and 50 mg test item/L, respectively. The results for nominal loadings of 25, 12.5 and 6.25 mg Carbon/L were below the Limit of Quantification, however, they were clearly below the NOEL.

The NOEL and LOEL after 96 hours of exposure were determined at 50 and 100 mg/L loading rate WAF respectively.

The 96-hour LC50 was determined to be > 100 mg/L loading rate WAF.