[No public or meaningful name is available]

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:

03 December 2014 - 05 December 2014

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Not GLP Compliant

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: ISO 14669: (Water Quality - Determination of acute lethal toxicity to marine copepods)

Version / remarks:

The study procedures described in this report were based on the ISO International Standard 14669:
"Water quality - Determination of acute lethal toxicity to marine copepods (Copepoda, Crustacea);
First edition 1999-04-01.

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OECD 23

Version / remarks:

Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

GLP compliance:

no

Remarks:

Study not done for EU Reach as an intermediate under SCC but to evaluate the test substance for its ability to generate acute lethal toxic effects on marine copepod Tisbe battagliai.

Test material

Test material information

Constituent 1

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Remarks:

The purpose of the analytical phase was to determine the actual concentrations in samples taken from the test solutions used during the ecotoxicity test

Details on sampling:

The test samples were stored in the freezer (≤ -15°C). Storage stability of samples under these conditions was demonstrated in project 506881.
On the day of analysis, the test samples were defrosted at room temperature. The samples were diluted in a 3:1 (v:v) ratio with acetonitrile and analysed.

Test solutions

Vehicle:

yes

Details on test solutions:

5 μm filtered and UV treated natural marine water of the same origin as the water that was used for culturing the organisms. Water was obtained from Guernsey Sea Farms Ltd., Guernsey, United Kingdom and stored at 4ºC until test initiation.

Test organisms

Test organisms (species):

other aquatic crustacea: Tisbe battagliai

Study design

Test type:

static

Water media type:

saltwater

Remarks:

UV treated natural marine water

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

24 adn 48 hours

Test conditions

Test temperature:

18-22°C, constant within 2°C.

pH:

Between 6.0 and 8.5.

Dissolved oxygen:

8.5-9.5 mg O2/l

Salinity:

34%-36%

Nominal and measured concentrations:

Solutions containing 1.0, 10 and 100% WSF prepared at a loading rate of 100 mg/l

Details on test conditions:

5.5.2. Test procedure and conditions
Test duration: 48 hours
Test type Static
Test vessels 24-well plastic plates covered with a lid.
Volume of vessel 3 ml
Medium 5 μm filtered and UV treated natural marine water of the same origin as the water that was used for culturing the organisms. Water was obtained from Guernsey Sea Farms Ltd., Guernsey, United Kingdom and stored at 4ºC until test initiation.
Specifications: dissolved oxygen concentration >80% of the air saturation value
pH: 8.0
Salinity: 35‰.
Number of copepodids 20 each in the control and the highest concentration
10 each in the lower concentrations
Loading 5 per vessel containing 2.5 ml of test solution
Light 16 hours photoperiod daily
Temperature 18-22°C, constant within 2°C.
Oxygen concentration ≥ 4 mg/l at the end of the test.
Aeration No aeration of the test solutions.
pH Between 6.0 and 8.5.
Feeding No feeding
Introduction of copepodids Within 0.5 hour after preparation of the test solutions.

5.5.4. Measurements and recordings
Mortality At 24 hours and at 48 hours.
pH and dissolved oxygen At the beginning and at the end of the test, in the control and the highest concentrations (measurements were performed in separate vessels).
Temperature of medium At the beginning of the test, for the control and the highest concentration. In addition, temperature was measured continuously in a temperature control vessel.

5.6. Electronic data capture
Observations/measurements in the study were recorded electronically using the following programme(s):
- REES Centron Environmental Monitoring system version SQL 2.0 (REES Scientific, Trenton, NJ, USA): Temperature.

5.7. Interpretation

5.7.1. Data handling
The 24 and 48h-LCx values could not be determined because test substance appeared to be not toxic.

5.7.2. Acceptability of the test
1. In the control, no copepodids became immobilised.
2. The oxygen concentration at the end of the test was ≥4 mg/l in control and test vessels.
3. The sensitivity of the batch Tisbe battagliai tested was within the range described by the protocol.

5.8. List of deviations

5.8.1. List of protocol deviations
1. Sea water was filtered with 5 instead 1 μm filter.
Evaluation: No other organisms were present tin the medium during the test. Sensitivity of the batch was within the range.
2. In two replicates (1.0% WSF A and 100% WSF B) six instead of 5 tisbes were tested.
Evaluation: This had no effect on the results.

The study integrity was not adversely affected by the deviations.

Reference substance (positive control):

yes

Remarks:

Test medium containing 0.3 mg Zn1/l (positive control).

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

LC10

Effect conc.:

> 0.005 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 2

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 0.005 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LC10

Effect conc.:

> 0.005 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 4

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 0.005 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Samples taken from the highest concentration were analysed. The actual concentration was 0.0072 mg/l at the start of the test. The measured concentration decreased to 0.0031 mg/l (43% of initial) at the end of the exposure. The average exposure concentration was calculated to correspond to 0.0047 mg/l. It should be noted that measured values were obtained by either by extrapolation of the calibration curve or calculated using the mean response factor of the lowest calibration solution. Therefore, the average exposure concentration should be considered indicative.

Results with reference substance (positive control):

At the end of the test 50% mortality was observed in the reference control. Since the effect in this treatment should be between 20 and 80% it was concluded that the batch tisbe used for this test showed normal sensitivity to Zinc sulphate heptahydrate.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of the present study with Tisbe battagliai the 48h-LC10 and 48h-LC50 were >0.0047 mg/l, considered the maximum solubility in medium.