Reaction mass of copper digluconate, sodium sulphate and copper sulphate

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:

07. Aug. 2008 - 04. Jun. 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

OECD Guideline for Testing of Chemicals, No. 202, Daphnia sp., Acute Immobilisation Test, 2004.

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Directive 92/69/EEC

Version / remarks:

EU Commission Directive 92/69/EEC, C.2, Acute Toxicity for Daphnia, 1992.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Copper di-D-gluconate

EC Number:

208-408-2

EC Name:

Copper di-D-gluconate

Cas Number:

527-09-3

Molecular formula:

C12H22CuO14

IUPAC Name:

copper(2+) bis(2,3,4,5,6-pentahydroxyhexanoate)

Constituent 2

Reference substance name:

Sodium sulphate

EC Number:

231-820-9

EC Name:

Sodium sulphate

Cas Number:

7757-82-6

Molecular formula:

H2O4S.2Na

IUPAC Name:

disodium sulfate

Constituent 3

Reference substance name:

Copper sulphate

EC Number:

231-847-6

EC Name:

Copper sulphate

Cas Number:

7758-98-7

Molecular formula:

CuSO4

IUPAC Name:

copper(2+) ion sulfate

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

not specified

Details on sampling:

Dosage
A stock solution with a nominal concentration of 44 mg/L was prepared by completely dissolving 44.3 mg of test item in 1000 mL of test water using ultrasonic treatment for 10 minutes and intense stirring for 10 minutes at room temperature. This intensively mixed stock solution was used in a series of dilutions to prepare the test media of all test item concentrations. The test media were prepared just before introduction of the daphnids (i.e., start of the test).

See more information in the study report attach

Test solutions

Vehicle:

not specified

Details on test solutions:

Study Design
A static test without test medium renewal was performed. The following nominal concentrations of test item were tested: 0.10, 0.22, 0.46, 1.0 and 2.2 mg/L. Additionally, a control (test water without test item) was tested in parallel. The selection of the test concentrations was based on the results of range-finding tests (non-GLP).
For each treatment, 20 daphnids were used divided into four replicates of five daphnids each.
The volume of test solution provided for each daphnia was 10 mL. Thus, the requirement of the test guidelines for the minimum volume of 2 mL test medium per daphnia was fulfilled. The daphnids were randomly distributed to the test vessels at initiation of the test.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna Straus
- Strain/clone: clone 5 (young daphnids of the species Daphnia magna Straus)

Test system
The study was performed with young daphnids of the species Daphnia magna Straus. A clone of this species (defined by the supplier as clone 5) was originally supplied by the University of Sheffield / UK in 1992. Since that time, the clone has been bred at Harlan Laboratories in reconstituted water of the quality identical to the water quality used in the tests (in respect to pH, main ions, and total hardness) and under temperature and light conditions identical to those of
the tests (see below).
At the start of the test, the organisms used in the test were 6-24 hours old and were not first brood progeny.
For evaluation of the quality of the daphnia clone and the experimental conditions, potassium dichromate is tested as a positive control twice a year. The result of the latest positive control test in October 2008 (48-hour EC50: 0.70 mg/L, Harlan Laboratories Study C25711) indicated that the sensitivity of the test organisms was within the historical range of the Harlan Laboratories (48-hour EC50 from 1996 to 2008: 0.53-1.1 mg/L).
The test method and the test species are recommended by the international test guidelines.

Study design

Test type:

not specified

Water media type:

not specified

Total exposure duration:

48 h

Test conditions

Hardness:

Not specified

Test temperature:

The water temperature was maintained at 20 °C. Specified in table 3 ( page 18 of the study report).

pH:

Specified in table 3 (page 18 of the study report).

Dissolved oxygen:

Specified in table 3 (page 18 of the study report).

Salinity:

Not specified

Conductivity:

Not specified

Nominal and measured concentrations:

Specified in table 3 (page 18 of the study report).

Details on test conditions:

Experimental Conditions
The test was performed in a temperature-controlled room with continuous monitoring of the room temperature. The water temperature was maintained at 20 °C.
A 16-hour light to 8-hour dark cycle with a 30-minute transition period was used. Light intensity during the light period was approximately between 520 and 680 Lux.
The daphnids were not fed during the test.

Reference substance (positive control):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.59 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

0.22 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

Effect concentrations 3

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.22 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

2.2 mg/L

Nominal / measured:

not specified

Conc. based on:

not specified

Basis for effect:

not specified

Details on results:

RESULTS AND DISCUSSION
No analytical work was performed in agreement with the Sponsor.

The biological results (based on nominal test concentrations) are listed in Table 1.

During the first 24 hours of the test, no immobilized test organisms were determined in the
control and up to and including the test item concentration of 0.22 mg/L. At the next higher
concentrations of 0.46, 1.0 and 2.2 mg/L, the immobilization was 15, 40 and 90%, respectively.

The 24-hour EC50 of the test item was calculated to be 1.0 mg/L with 95% confidence limits of
0.36 and 3.1 mg/L. The 24-hour EC0 was 0.22 mg/L. The 24-hour EC100 was >2.2 mg/L.

After 48 hours of exposure, no immobilized test organisms were determined in the control and up to and including the test item concentration of 0.22 mg/L. At the next higher concentrations of 0.46 and 1.0 mg/L, the immobilization increased to 35 and 85%, respectively. At the highest test concentration of 2.2 mg/L, all test organisms were found to be immobile.

The 48-hour EC50 was calculated to be 0.59 mg/L with 95% confidence limits of 0.25 and
1.4 mg/L. The 48-hour EC0 and the 48-hour NOEC (highest concentration tested without toxic effects after 48 hours) of test item were both 0.22 mg/L, since no immobilization was observed up to and including this test concentration. The 48-hour EC100 was 2.2 mg/L.
No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions throughout the entire test duration (Table 2).

At the beginning and end of the test period, the dissolved oxygen concentrations in the test media and control were at least 8.2 mg/L, the pH value of the test media was in the range of 7.8 to 8.0, and the water temperature during the test was 20 °C (Table 3).

Results with reference substance (positive control):

The biological test results (based on nominal test concentrations) were as follows:

- 24-hour EC50: 1.0 mg/L
(95% confidence limits: 0.36 and 3.1 mg/L)
- 24-hour EC0: 0.22 mg/L
- 24-hour EC100: >2.2 mg/L
- 48-hour EC50: 0.59 mg/L
(95% confidence limits: 0.25 and 1.4 mg/L)
- 48-hour EC0 and 48-hour NOEC: 0.22 mg/L
- 48-hour EC100: 2.2 mg/L

Any other information on results incl. tables

See the tables in the study report from page 16 to page 18

Table 1 Effect of Sergomil L60 on the Mobility of Daphnia magna

Table 2 Appearance of the Test Media during the Test Period

Table 3 Dissolved Oxygen Concentrations, pH Values, and Temperature in the Treatments

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

EC50 (48h) =0.59 mg/L (<1mg/L). Substance classifies as Aq. Acute tox 1.

Executive summary:

The acute toxicity of the test item to Daphnia magna was determined in a 48-hour static test according to the EU Commission Directive 92/69/EEC, Part C.2 and the OECD Guideline for Testing of Chemicals, No. 202 (2004).

The nominal test item concentrations tested were 0.10, 0.22, 0.46, 1.0 and 2.2 mg/L. Additionally, a control group was tested in parallel.

No analytical work was performed in agreement with the Sponsor.

The biological test results (based on nominal test concentrations) were as follows:

- 24-hour EC50: 1.0 mg/L

(95% confidence limits: 0.36 and 3.1 mg/L)

- 24-hour EC0: 0.22 mg/L

- 24-hour EC100: >2.2 mg/L

- 48-hour EC50: 0.59 mg/L

(95% confidence limits: 0.25 and 1.4 mg/L)

- 48-hour EC0 and 48-hour NOEC: 0.22 mg/L

- 48-hour EC100: 2.2 mg/L