Iron(1+), chloro[dimethyl 9,9-dihydroxy-3-methyl-2,4-di(2-pyridinyl-kN)-7-[(2-pyridinyl-kN)methyl]-3,7-diazabicyclo[3.3.1]nonane-1,5-dicarboxylate-kN3,kN7]-, chloride

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:

From 2004-02-09 to 2004-06-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,5-dimethyl 3-methyl-9-oxo-2,4-bis(pyridin-2-yl)-7-[(pyridin-2-yl)methyl]-3,7-diazabicyclo[3.3.1]nonane-1,5-dicarboxylate dichloroiron hydrate

EC Number:

610-388-9

Cas Number:

478945-46-9

Molecular formula:

C28H31N5O6FeCl2

IUPAC Name:

1,5-dimethyl 3-methyl-9-oxo-2,4-bis(pyridin-2-yl)-7-[(pyridin-2-yl)methyl]-3,7-diazabicyclo[3.3.1]nonane-1,5-dicarboxylate dichloroiron hydrate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: S2539801
- Expiration date of the lot/batch:
- Purity test date: 2004-11-12

RADIOLABELLING INFORMATION (if applicable) no

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
ambient/dark

TREATMENT OF TEST MATERIAL PRIOR TO TESTING: no

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0-10-18-32-56-100 mg/L
- Sampling method: analysis was carried out at 0H, 24H (old and new) and 48H to assess the satisfactory achieved concentration and stability of the test concentrations of 0, 10, 18, 32, 56 and 100 mg/L
- Sample storage conditions before analysis: not stated.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a nominal 1000 mg/L stock was prepared by dissolving 1000 mg of test item in 1000 mL of purified water. test media were prepared by diluting the stock with appropriate volumes of Elendt M7 medium to obtain nominal concentrations of 0, 10, 18, 32, 56 and 100 mg/L. 12 L of Elendt M7 media were isolated and adjusted to pH7.5 using 1M HCl for use as dilution media to ensure that the pH of the test solutions remained below pH8
- Controls:Elendt M7 media
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): not reported.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: not stated
- Source: not stated
- Age of parental stock (mean and range, SD): not stated; juveniles <24H were used for the test
- Feeding during test: no

ACCLIMATION not stated

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

206 +/-14 mg/L as caCO3

Test temperature:

19.7+/-0.4°C

pH:

7.5+/-0.3

Dissolved oxygen:

8.0+/-0.1 mg/L

Nominal and measured concentrations:

Nominal: 0-10-18-32-56-100 mg/L
Mean measured: <3-7.30-13.49-23.19-42.93-79.42 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 120 mL crystallising dish, filled with approximately 100 mL tets media
- Aeration: not stated
- Renewal rate of test solution (frequency/flow rate): complete renewal at 24H
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates):4
- No. of vessels per control (replicates):4
- Biomass loading rate: 5 daphnia in 100 mL

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M7
- Culture medium different from test medium: not stated
- Intervals of water quality measurement: pH, dissolved oxygen and total hardness of the test media were measured at 0, 24 (old and new) and 48 hours. The temperature in a representative sample of the test vessels was measured daily.

OTHER TEST CONDITIONS
- Adjustment of pH:no
- Photoperiod: 16/8
- Light intensity: normal laboratory light intensity

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mobility at 24 and 48H

VEHICLE CONTROL PERFORMED: not appropriate

RANGE-FINDING STUDY
- Test concentrations: not reported
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

23.7 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

7.3 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Behavioural abnormalities: not reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: not reported
- Effect concentrations exceeding solubility of substance in test medium: no

Reported statistics and error estimates:

The EC50 was calculated using the computer application BMPDIN. the program generates where possible, three statistical analyses of the data using the moving average method, the probit method and the binomial method.

Any other information on results incl. tables

An anomalous result was observed in the 10 mg/L (24H new) test solution. the original test sample supplied to chemical analysis had a mean measured concentration of 17.42 mg/L (174.2% nominal). A second test sample of 10 mg/L (24H new) solution was supplied and the original sample re-analysed. The mean measured concentration of the re-analysed test sample was 15.38 mg/L (153.8% nominal) and the concentration of the second sample was 8.06 mg/L (80.6% nominal). Re-analysis of the original sample confirmed that loss of the test compound over time was not significant and suggested that this was not the reason for the lower concentration in the second sample. It was believed that the original sample supplied may have been taken from the wrong test solution flask as the results would appear to correlate more closely with the 18 mg/L test solution. Consequently a mean measured concentration of 8.06 has been taken as being the correct and used in the calculation of the results.

Effect data:

Mean measured conc. (mg/L) Number immobile at 24H  Number immobile at 48H        Number mobile at 48H                  Replicate             Replicate             Replicate      1 2   3  4  total  1  2  3  4  Total  1  2 3   4  Total  0.0  0  0  0  0  0  0  0  0  0  0  5  5  5  5  20  7.3  0  0  0  0  0  0  0  0  0  0  5  5  5  5  20  13.5  0  0  0  0  0  1  0  1  0  2  4  5  4  5  18  23.2  0  0  1  1  2  2  3  0  0  7  3  2  4  4  13  42.9  3  3  3  4  13  2  2  2  1  20  0  0  0  0  0  79.4  5  5  5  5  20  -  -  -  -  20  0  0  0  0  0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

immobility in control: 0% at 48H; dissolved oxygen: 8.0+/-0.1 mg/L in all vessels

Conclusions:

48H-EC50=23.7 mg/L based on mean measured concentrations.
48H-NOEC=7.3 mg/L based on mean measured concentrations.

Executive summary:

The purpose of the study was to determined the 48H EC50 of the test item to juvenile <24 old Daphnia magna, following to OECD 202 guideline and C2 method and according to GLP. The test was conducted under semi-static conditions. Daphnia magna were not fed during the test. Daphnia were exposed to a range of concentrations of the test item prepared by diluting a concentrated stock solution with Elendt M7 medium. Nominal test concentrations were 0-10-18 32-56-100 mg/L and were determined by liquid chromatography/mass spectrometry. Mean measured test concentrations were <3 -7.30 -13.49 -23.19 -42.93 -79.42 mg/L.

48H-EC50=23.7 mg/L based on mean measured concentrations.

48H-NOEC=7.3 mg/L based on mean measured concentrations.

All validity criteria were fulfilled.