1,3-Diazaspiro[4.5]decane-2,4-dione, 8-methoxy-, sodium salt (1:1), cis-

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:

2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

sodium (5s,8s)-8-methoxy-2,4-dioxo-1,3-diazaspiro[4.5]decan-3-ide

EC Number:

929-513-6

Cas Number:

1400584-86-2

Molecular formula:

C9H13N2O3 · Na

IUPAC Name:

sodium (5s,8s)-8-methoxy-2,4-dioxo-1,3-diazaspiro[4.5]decan-3-ide

Test material form:

solid: bulk

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

49.0 mg of the test item were dissolved in Milliporewater and made up with Milliporewater to the mark in a 100 mL volumetric flask to prepare a stock solution of 490 mg/L. Defined volumes of this stock solution were diluted with Milliporewater to obtain standard solutions in the range of 0.025 to 4.90 mg/L.

Sample storage conditions before analysis: Routinely, the samples are analysed immediately. Only in exceptional cases, they are stored overnight deeply frozen and protected from light.

Test solutions

Vehicle:

no

Details on test solutions:

To produce the only test concentration 105.6 mg of the test item were added to 1 litre of dilution water, treated for 10 minutes in an ultrasonic bath and then stirred for 10 minutes on a magnetic stirrer. The pH was measured to be pH 9.2 and was adjusted to pH 8.0.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Common name: water flea
Source: Strain of Bundesgesundheitsamt Berlin
Maintenance and Acclimatisation:
A population of parthenogenetic females of synchronized age structure is maintained since more than 15 years in the test facility under constant temperature conditions (20 +/- 1 °C) at a 16 : 8 hour light-dark photoperiod (illumination: < 1000 lux).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

The hardness of the dilution water used was 14.8 °dH (= 264 mg/L CaCO3).

Test temperature:

20.5 - 20.7 °C

pH:

7.7

Dissolved oxygen:

8.7 mg/L

Nominal and measured concentrations:

nominal: 100 mg/L
measured: 103.7 mg/L (0 h), 97.1 mg/L (48 h)

Details on test conditions:

Test vessels: 50 mL glass beakers holding 10 neonates in 20 mL of test medium

Experimental design:
1 test concentration plus 1 control
10 neonates per vessel, 2 replicates per concentration/control
no feeding during the exposure period

Photoperiod: 16 h light: 8 h dark

Aeration: none

Criteria of effects:
The criterion of adverse effects used in this study was the item-induced alteration of the normal mobility behaviour and the loss of locomotory actions of the neonates, observed at 24 and 48 hours.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

No toxic effects against daphnia at a limit test concentration of 100 mg/L.

Executive summary:

A study was performed to assess the acute toxicity to Daphnia magna STRAUS under static conditions.
The study was conducted in accordance with the OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test' (adopted April 13, 2004) which is in most parts equivalent to the EEC Methods for Determination of Ecotoxicity Annex to Directive 92/69/EEC (OJ. No. L383A, 29.12.92) Part C, Method 2 Acute toxicity for Daphnia'.
The Daphnia were exposed to a limit test concentration of nominally 100 mg/L of the test item dissolved in water. Auxiliaries used to prepare the test media were an ultrasonic bath and a magnetic stirrer.
Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. The following values were determined:

Time  [h]	EC 50 [mg/L]
24	> 100
48	> 100

No toxic effects against daphnia at a limit test concentration of 100 mg/L.
The results are expressed in terms of nominal concentrations. Recovery rates correspond to 103.7% of nominal values at 0 hours, and to 97.1% of nominal values at 48 hours, respectively.