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

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:

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 203 (Fish, Acute Toxicity Test)

Version / remarks:

2019

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

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:

Sampling schedule:
Control: at 0 and 96 hours
Test concentration: at 0 and 96 hours

Test solutions

Vehicle:

no

Details on test solutions:

To achieve the desired test concentration 502.0 mg of the test item were added to 5 litres of dilution water.
The pH was to be pH 9.2 and was adjusted to pH 7.9.
The stock solution was directly used to prepare the exposure concentration.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Name: Zebra fish
Source: Aqua KlöGer (Germany)
Date supplied: 2007-02-13
Acclimatisation: Stock held since 2007-02-13 and acclimatised to the test conditions since then.
Temperature: 20 - 24 °C
Dissolved oxygen: > 5 mg/L
Feeding: Commercial fish food, daily. Discontinued 24 h prior to test.
Mortalities during acclimatisation period: < 5 %
Medication: none

Mean standard length (n = 20): 3.5 cm (S.D. = 0.20 cm)

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

The total hardness of the dilution water, measured at test start, was 15.5 °dH (= 277 mg/L CaCO3).

Test temperature:

21.0 - 22.6 °C

pH:

7.7 - 8.1

Dissolved oxygen:

8.1 - 8.5 mg/L

Nominal and measured concentrations:

nominal: 100 mg/L
measured: 99.6 mg/L (0 and 96 hours)

Details on test conditions:

Reconstituted water prepared according to the recommendations of ISO 7346. This freshly prepared standard dilution water is used for both, the maintenance of the test animals under flow-through conditions and the preparation of stock and test solutions of the test item.

Exposure conditions
Test vessels: glass aquaria holding 5 L of test media

Experimental design:
1 test concentration plus 1 control
10 animals per test concentration

Loading: 0.74 g body weight (wet weight) per litre
Photoperiod: 16 h light: 8 h dark
Aeration: gentle aeration via narrow glass tubes

Criteria of effects:
The criterion of death used in this study was the absence of response to physical stimulation.
In addition to observations on mortality at 2, 24, 48, 72 and 96 hours, subjective assessments were made on the type and incidence of sub-lethal effects compared with control fish.

Reference substance (positive control):

not required

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

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

Executive summary:

A study was performed to assess the acute toxicity of the test item to Danio rerio under static conditions.
The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity Annex to Directive 92/69/EEC (OJ. No. L383A, 29.12.92) Part C, Method 1 'Acute toxicity for Fish' which is in most parts equivalent to the OECD Guideline for Testing of Chemicals No. 203 'Fish, Acute Toxicity Test'.
Groups of ten fish of the recommended size were exposed to a limit test concentration of nominally 100 mg/L test item dissolved in water.
Observations were made on the number of dead fish and the incidence of sub-lethal effects after 2, 24, 48, 72 and 96 hours of exposure. The following values were determined:

Time [h]	LC 50 [mg/L]
2	> 100
24	> 100
48	> 100
72	> 100
96	> 100

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

All results are expressed in terms of nominal concentrations. Recovery rates correspond to 99.6% of nominal values at 0 and 96 hours, respectively.