Aluminium Bordeaux

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:

2017-04-10 to 2017-05-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

OECD Guideline; EU Method; GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

disodium 2,4-dihydroxy-5-[(2-hydroxy-3-nitro-5-sulphonato-phenyl)azo]benzoate, reaction products with chromium triacetate

Molecular formula:

not applicable

IUPAC Name:

disodium 2,4-dihydroxy-5-[(2-hydroxy-3-nitro-5-sulphonato-phenyl)azo]benzoate, reaction products with chromium triacetate

Sampling and analysis

Analytical monitoring:

no

Remarks:

No analytical dose verification was performed since the test substance (type UVCB) cannot be analysed by standard methods in such low test concentrations used in this study.

Test solutions

Vehicle:

no

Details on test solutions:

The test medium of the only test item concentration of nominal 100 mg/L was prepared by dissolving 40.0 mg test item into 400 mL test water by intense stirring for 23.75 hours and ultrasonic treatment for 10 minutes.
The test media was prepared just before introduction of the daphnids (= start of the test).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Straus), clone 5
Age at Test Start: From 5.25 to 22.5 hours old
Sex: Female
Origin: The daphnids introduced in the test were taken from ibacon's in house laboratory culture.
Breeding Conditions: The daphnids were bred in the laboratories of ibacon under similar temperature and light conditions as used in the test. The cultivation of the parental daphnids was performed in Elendt M4 medium. The test organisms were not first brood progeny. The daphnids in the stock culture were fed at least on all working days with green algae (Desmodesmus subspicatus) freshly grown in the laboratories of ibacon.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Water Hardness: 2.5 mmol/L (= 250 mg/L) as Calcium Carbonate

Test temperature:

20.6 to 21.4 °C at test start;
20.1 to 20.3 °C at test end

pH:

7.6 to 8.1 at test start;
7.7 to 8.0 at test end; and thus the pH-value did not vary by more than 1.5 units

Dissolved oxygen:

8.3 to 8.7 mg/L at test start;
8.3 to 8.6 mg/L at test end

Nominal and measured concentrations:

100 mg test item/L and a control

Details on test conditions:

Test Environment: Controlled environment room
Measurement of pH, Dissolved
Oxygen and Water Temperature:
The water temperature, pH-values and dissolved oxygen concentrations were determined at test start and test end in each treatment group.
Water Temperature: 20.6 to 21.4 °C at test start;
20.1 to 20.3 °C at test end
pH-Values: 7.6 to 8.1 at test start;
7.7 to 8.0 at test end; and thus the pH-value did not vary by more than 1.5 units
Dissolved Oxygen Concentration: 8.3 to 8.7 mg/L at test start;
8.3 to 8.6 mg/L at test end
Light Regime: 16 h light : 8 h dark
Light Intensity: The light intensity was 540 to 570 lux (measured once during the test).

Reference substance (positive control):

yes

Remarks:

For the evaluation of the quality of the Daphnia clone and the consistency of the experimental conditions, the reference item potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

After 48 hours of exposure, no immobilisation of the test animals was observed in the control and in the only test concentration of nominal 100 mg test item/L.

Results with reference substance (positive control):

In the most recent test with the reference item potassium dichromate performed in January 2017 the EC50 after 24 hours was determined to be 1.48 mg test item/L, indicating that the sensitivity of the daphnids was consistent with the level proposed by the OECD 202 guideline (EC50-24 h between 0.6 and 2.1 mg potassium dichromate/L).

Reported statistics and error estimates:

No statistical analysis was performed.
The EC50 could not be quantified due to the absence of toxicity of the test item. The NOEC and the LOEC were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Control Immobilisation Rate: 0 % and furthermore no daphnid showed signs of disease or stress. Dissolved Oxygen Concentration: >= 8.3 mg O2/L in the control and test vessels at the end of the test.

Conclusions:

The toxic effect of the test item to Daphnia magna was assessed in a static limit test. The 48-hour NOEC was determined to be at least 100 mg test item/L. The NOEC might even be higher than this concentration, but concentrations in excess of nominal 100 mg test item/L have not been tested according to the recommendations of the test guidelines. The 48-hour LOEC and the 48-hour EC50 were clearly higher than 100 mg test item/L. These values could not be quantified due to the absence of toxicity of the test item up to nominal 100 mg test item/L. No analytical dose verification was performed.

Executive summary:

Title:     Acute Toxicity to Daphnia magna in a Static 48-hour Immobilisation Limit Test

Purpose:             The purpose of this study was to determine the influence of the test item on the mobility of Daphnia magna.

For this purpose, young daphnids (< 24 hours old) were exposed in a static test for 48 hours to the only concentration of nominal 100 mg/L under defined conditions for 48 hours. The recorded effects were the mobility of the daphnids after 24 and 48 hours.

This limit-test was performed in compliance with the test guidelines in order to demonstrate that the test item has no toxic effects on the daphnids up to this concentration.

The test method of application and the test system are recommended by the test guidelines and Daphnia magna is the recommended test species.

No analytical dose verification was performed since the test substance (type UVCB) cannot be analysed by standard methods in such low test concentrations used in this study.

Guidelines:        Commission Regulation (EC) No 440/2008, Annex, Part C, C.2.: "Daphnia sp. Acute Immobilisation Test", Official Journal of the European Union (EN), dated May 30, 2008;             OECD Guideline for Testing of Chemicals No. 202: "Daphnia sp., Acute Immobilisation Test" adopted April 13, 2004              

Material and Methods  

Test Item:          batch no.: Op 01/15; type: UVCB substance, according to certificate of analysis

Test Species:     Female Daphnia magna, clone 5; 5.25 to 22.5 hours old

Source: The daphnids introduced in the test were taken from ibacon's in-house laboratory culture.

Test Design:     This study encompassed two treatment groups (one test item concentration at nominal 100 mg/L, and one control) each containing 20 individuals. The mobility of the daphnids was determined in a static 48-hour test by visual observation after 24 and 48 hours.

Endpoints:         Number of immobile organisms after 24 and 48 hours

Test Concentrations:     100 mg test item/L and a control.

Test Conditions:              Water temperature: 20.1 to 21.4 °C; pH value: 7.6 to 8.1; dissolved oxygen concentration: 8.3 to 8.7 mg/L; photoperiod: 16 h light - 8 h dark; light intensity: 540 to 570 lux; and thus were within the ranges requested by guideline OECD 202

Results

Biological test results:   After 48 hours of exposure, no immobilisation of the test animals was observed in the control and in the only test concentration of nominal 100 mg test item/L.

Conclusion:        The toxic effect of the test item to Daphnia magna was assessed in a static limit test. The 48-hour NOEC was determined to be at least 100 mg test item/L. The NOEC might even be higher than this concentration, but concentrations in excess of nominal 100 mg test item/L have not been tested according to the recommendations of the test guidelines. The 48-hour LOEC and the 48-hour EC50 were clearly higher than 100 mg test item/L. These values could not be quantified due to the absence of toxicity of the test item up to nominal 100 mg test item/L. No analytical dose verification was performed.

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.