Reaction mass of copper digluconate, sodium sulphate and copper sulphate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07.Ag.2008 - 04.Apr. 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 203 (Fish, Acute Toxicity Test)

Version / remarks:

OECD Guideline for the Testing of Chemicals, No. 203, Fish, Acute Toxicity Test, 1992.

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: Directive 92/69/EEC

Version / remarks:

EU Commission Directive 92/69/EEC, C.1: Acute Toxicity for Fish, 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 concentrated stock solution of nominal 500 mg/L was daily prepared by completely dissolving test item in test water using ultrasonic treatment for 10 minutes and stirring for 10 minutes at room temperature. The following test item amounts were weighed into the test water:
Weighed amount of test item (mg) Amount of test water (mL)
2251.2 4500
750.3 1500
99.9 200
99.8 200

Adequate volumes of this intensively stirred stock solution were added to the test water in the test vessels and were intensively mixed to prepare the test media with the test concentrations as stated above.
The test media were daily freshly prepared just before introduction of the fish (= start of the test) and before each test medium renewal.

Test solutions

Vehicle:

not specified

Details on test solutions:

Study Design
The following nominal concentrations of Sergomil L60 were tested: 0.32, 1.0, 3.2, 10, 32 and 100 mg/L. Additionally, a control (test water without test item) was tested in parallel.
A semi-static test procedure was chosen. The test media were renewed daily to keep the concentrations of Sergomil L60 in the test media as constant as possible during the test period of 96 hours.
During this semi-static test, the test fish were daily placed into a new test vessel with freshly prepared test medium of the corresponding treatment.
The selection of the test concentrations was based on the results of a range-finding test (non- GLP).
At the start of the test, 7 fish were introduced into each test vessel in a random order. The loading rate was 0.64 g fish wet weight per liter test medium. Thus, the requirement of a loading rate of less than 1 g fish/L was fulfilled.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Strain: Oncorhynchus mykiss
- Source:
- Age at study initiation (mean and range, SD): juvenile
- Length at study initiation (length definition, mean, range and SD):
- Weight at study initiation (mean and range, SD):
- Method of breeding:
- Maintenance of the brood fish:

ACCLIMATION
- Acclimation period:
- Acclimation conditions (same as test or not):
- Type and amount of food during acclimation:
- Feeding frequency during acclimation:
- Health during acclimation (any mortality observed):

QUARANTINE (wild caught)
- Duration:
- Health/mortality:

FEEDING DURING TEST
- Food type:
- Amount:
- Frequency:

Study design

Test type:

semi-static

Water media type:

other: Local tap water (non chlorinated well water of drinking water quality)

Total exposure duration:

96 h

Test conditions

Hardness:

The total hardness of the test water was 214 mg/L as CaCO3.

Test temperature:

The water temperature in the test vessels was maintained at 14-15 °C (Table 5).

pH:

The pH values in the test media and the control ranged from 8.0 to 8.4 (Table 3).

Dissolved oxygen:

The oxygen concentration was always 9.1 mg/L or higher (Table 4), and thus higher than 60% oxygen saturation.

Salinity:

Not specify

Conductivity:

Not specify

Nominal and measured concentrations:

The nominal test item concentrations tested were 0.32, 1.0, 3.2, 10, 32 and 100 mg/L. Additionally, a control group was tested in parallel.

Details on test conditions:

Experimental Conditions
The water temperature in the test vessels was maintained at 14-15 °C (Table 5). The test vessels were slightly aerated during the test period.
A 16-hour light to 8-hour dark photoperiod, with a 30-minute transition period was used (light intensity during the light period was approximately within the range of 100 to 600 Lux).
The test duration was 96 hours and the fish were not fed during the test.

Reference substance (positive control):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

not specified

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

not specified

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

not specified

Effect concentrations 4

Key result

Duration:

96 h

Dose descriptor:

LC50

Remarks:

95% confidence interval of 1.0-3.2 mg/L.

Effect conc.:

1.8 mg/L

Nominal / measured:

nominal

Conc. based on:

not specified

Basis for effect:

not specified

Effect concentrations 5

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

3.2 mg/L

Nominal / measured:

nominal

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 reported biological results were based on the nominal concentrations of the test item.

The biological results (observations for visible abnormalities in the test fish, mortalities and the LC50 values) are listed in Table 1. In the control and at the test concentrations up to and including 1.0 mg/L, all fish survived until the end of the test and no visible abnormalities were observed in the test fish. At all higher test concentrations, all fish were dead already after 24 to 48 hours of test duration.

Thus, the 96-hour NOEC and the 96-hour LC0 of test item to rainbow trout were both determined to be 1.0 mg/L. The 96-hour LOEC was 3.2 mg/L.

The 96-hour LC50 of test item was calculated to be 1.8 mg/L with a 95% confidence interval of 1.0-3.2 mg/L. The 96-hour LC100 was 3.2 mg/L. Thus, the concentration-effect relationship of the test item was rather steep.
No remarkable observations were made concerning the appearance of the test media. All test media were clear solutions colored by the test item at the highest test concentration of 100 mg/L throughout the entire test duration (Table 2).

The pH values in the test media and the control ranged from 8.0 to 8.4 (Table 3). The oxygen concentration was always 9.1 mg/L or higher (Table 4), and thus higher than 60% oxygen saturation. The water temperature was 14-15 °C during the test (Table 5).

Results with reference substance (positive control):

The biological test results after 96 hours of test duration:
– 96-hour LC50: 1.8 mg/L
(95% confidence interval: 1.0-3.2 mg/L)
– 96-hour LC0: 1.0 mg/L
– 96-hour LC100: 3.2 mg/L
– 96-hour NOEC: 1.0 mg/L
– 96-hour LOEC: 3.2 mg/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Substance presents no acute toxicity for fish in 96h.

Executive summary:

The acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss) was determined in a 96-hour semi-static test with a daily test medium renewal according to the EU Commission Directive 92/69/EEC, Part C.1 and the OECD Guideline for Testing of Chemicals No. 203, (1992).

The nominal test item concentrations tested were 0.32, 1.0, 3.2, 10, 32 and 100 mg/L. Additionally, a control group was tested in parallel.

No analytical work was performed in agreement with the Sponsor. The reported biological results were based on the nominal concentrations of the test item.

The biological test results after 96 hours of test duration:

– 96-hour LC50: 1.8 mg/L

(95% confidence interval: 1.0-3.2 mg/L)

– 96-hour LC0: 1.0 mg/L

– 96-hour LC100: 3.2 mg/L

– 96-hour NOEC: 1.0 mg/L

– 96-hour LOEC: 3.2 mg/L