Plum, ext.

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 August 2018 - 23 August 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Plum, ext.

EC Number:

290-179-3

EC Name:

Plum, ext.

Cas Number:

90082-87-4

Molecular formula:

not applicable

IUPAC Name:

Extract obtained from fruits of Prunus domestica L. Rosaceae

Test material form:

liquid

Details on test material:

- Physical appearance: blackish brown to dark brown liquid
- Storage conditions: at room temperature

Specific details on test material used for the study:

- No correction factor for purity is required and therefore not applied
- The test item is soluble and stable in water

Sampling and analysis

Analytical monitoring:

yes

Remarks:

TOC analysis

Details on sampling:

Single samples for possible analysis were taken from all concentrations and the control according to the schedule below.
Frequency: at t=0 h and t=48 h
Volume: 40 mL
Storage: Samples were stored in a refrigerator (2-8°C) until analysis.
At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation of test solutions started with the highest concentration of 100 mg/L applying a 15- minute period of magnetic stirring to accelerate the dissolution of the test item in test medium. Lower test concentrations were prepared by subsequent dilutions of the highest concentration in test medium.
- Evidence of undissolved test material: the highest test concentration was clear and of very slight yellow coloration, while the lower test concentrations were clear and colourless.
- Controls: Test medium without test item or other additives

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus, 1820
- Source: in-house laboratory culture with a known history. At least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
- Age of parental stock: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
- Age at study initiation: < 24 hours, from parental daphnids of more than two weeks old.
- Feeding during test: no

ACCLIMATION
- Acclimation period: no

BREEDING:
- Method: each batch was started with newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium (M7) in an all-glass culture vessel. Cultures were max. 4 weeks old. After 7 days of cultivation half of the medium was renewed twice a week.
- Feeding: daily, a suspension of fresh water algae.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg/L

Test temperature:

20-21°C

pH:

Control: 7.9 (t=0 h) and 8.0 (t=48 h)
100 mg/L: 7.8 (t=0 h and t=48 h

Dissolved oxygen:

Control: 8.5 mg/L (t=0 h) and 8.7 mg/L (t=48 h)
100 mg/L: 8.4 mg/L (t=0 h and t=48 h)

Nominal and measured concentrations:

Nominal concentration: 100 mg/L
Nominal TOC concentration: 32 mg/L
Measured TOC concentration: at t=0: 26.4 mg/L; at t=48: 21.8 mg/L (83% relative to nominal at t=0 h)

Details on test conditions:

TEST SYSTEM
- Test vessel: 60 mL, all-glass
- Fill volume: 50 mL
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 for 100 mg/L; 2 for 0.10, 1.0 and 10 mg/L
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Standard medium used: yes, according to OECD 202
- Source/preparation of dilution water: tap water purified by reverse osmosis
- Culture medium different from test medium: yes, M7 was used as culture medium

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: daily photoperiod of 16 hours

EFFECT PARAMETERS MEASURED: mobility at 24 and 48 hours.
- Additional measurements: pH and dissolved oxygen at the beginning and at the end of the test, in the limit test concentration and the control. Temperature of the medium: continuously in a temperature control vessel.

COMBINED LIMIT/RANGE-FINDING STUDY: yes, twenty daphnids per concentration (four replicates, 5 daphnids per vessel) were exposed to a control and test concentrations of 0.10, 1.0, 10 and 100 mg/L. No immobility was observed and the results were interpreted as limit test for determination of the effect concentrations.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (July 2018)

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Results of TOC analysis: the TOC of the test item was 31.78%, this value was used to estimate the nominal TOC concentration for the limit concentration of 100 mg/L. At the start of the test, 83% of the nominal TOC concentration were measured in the limit concentration, i.e. 26 mg/L. At the end of the test, the TOC concentration decreased slightly and was at 83% relative to the initial concentration, i.e. was 22 mg/L. Since TOC-analysis is a non-specific method, the effect parameters were determined based on the nominal concentration.

Results of the combined limit/range-finding study: no immobility was observed in the control group or at any of the test concentrations in the 48 hours exposure period.

- No biological, behavioural or other abnormalities were observed in the control group or any of the test concentrations.
- Individual pH, temperature and dissolved oxygen values remained within acceptable limits throughout the duration of the study.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Test concentrations: 0, 0.10, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/L
- 48h-EC50: 0.59 mg/L, 95%-confidence interval: 0.53-0.72 mg/L
- Other: results fell within the historical range of the test facility.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

1). In the control, no daphnids became immobilised or showed other signs of disease or stress. 2). The oxygen concentration at the end of the test was ≥ 3 mg/L in control and test vessels (i.e., 8.7 mg/L).

Conclusions:

The 48h-EC50 of Plum concentrate towards Daphnia magna, was determined to exceed the highest tested concentration (> 100 mg/L).