Perhydroimidazo[4,5-d]imidazole-2,5-dione

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPP 72-1 (Fish Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch Number: Fixapret 140 Nr. 766.
Date of Manufacturing: March 24 2001.
Purity: 96.3g/100g (determined by HPLC analysis).

Analytical monitoring:

yes

Details on sampling:

An analytical method for the determination of the test substance in the test water was developed by the Bioanalytical Laboratory of the testing facility. Samples for analytical determination of the concentration were taken within the first hour after the start of exposure and at the end of the exposure (approximately after 96 hours).

Concentration control analysis
Vehicle: Tap water.
Storage conditions of the samples until analysis: ambient or in refrigerator.
Sample preparation for analysis: Aliquots of the samples were directly used for HPLC-analysis.

Test water samples were collected from the middle section of the test vessel using a glass pipette and decanted in glass ampoules (which was rinsed with the test solution before they were filled). The transport to the Analytical Laboratory was done on the day of sampling.

Vehicle:

yes

Remarks:

Tap Water

Details on test solutions:

The dissolvation of the test substance in water is slow. Subsequently, the test solutions were therefore prepared a day prior to the start of the exposure, even after this there were still some residual undissolved material at the bottom of the test vessel; a wing stirrer was inserted for three hours to ensure complete dissolvation before the test organisms were introduced.

All test batches were present as clear solutions over the exposure period according to visual inspection.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ANIMALS
Supplier: Forellenzucht Trostadt GbR, Dorfstrasse 7, 98646 Trostadt, Germany (all animals used for the study were derived from the same batch of fish).
Body Weight (wet weight): 2.1g (1.5 -2.7g).
Body Length (from top of the snout to the end of the caudal fin): 6.1 cm (5.2 -6.5 cm).
Age: Approx. 5 months.
Physical Conditions: No signs of sickenss, injuries or abnormalities were noted.

ENVIRONMENTAL CONDITIONS
Acclimatisation period: 14 days.
Photoperiod: 16 hours light / 8 hours dark.
Water Quality: Non chlorinated charcoal filtered tap water (Frankenthal, Germany)
- Total hardness: Approx. 2.5 mmol/L = 250 mg/L CaCO3.
- Acid capacity: 5.5 mmol/L.
- Oxygen content: >80% saturation.
- pH value: Generally 7.5-8.5.
- Temperature: Approx. 12°C.
Diet: Growing feed (Forellenfutter (Zeigler), Supplier: Provimi Kuba AG, Gossau, Switzerland), ad libitum. Additionally (on workdays live or frozen shrimp brine (artemia) were provided.
Withdrawal of feed: During last day before start of exposure.
Medical treatment: None during acclimatisation.
Mortality during last week before start of exposure: 0.5%.

Test type:

static

Water media type:

other: Non-chlorinated charcoal filtered tap water (Frankenthal, Germany), aerated.

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

No mortality or clinical symptoms observed.

Hardness:

2.5 mmol/L = 250 mg/L CaCO3.

Test temperature:

12°C.

pH:

7.5 - 8.5.

Dissolved oxygen:

>80% saturation. Aeration was not performed over the first 72 hours. Slight aeration during the last day of exposure was carried out, since the oxygen content after 72 hours was decreased to 57% of the saturation value.

Salinity:

Not recorded.

Conductivity:

Approximately 550 µS/cm (25°C)

Nominal and measured concentrations:

Nominal test concentrations were 0mg/L and 100mg/L (weighed amount of test substance 5g in 50L of test water).

Details on test conditions:

Exposure was carried out in a static sysem.
Test vessel: Glass aquaria with stainless steel frame (60cm long, 35cm wide, 40cm high), water depth about 27cm.
Volume of water: 50L
Animals per test vessel: 10.
Loading (g fish/L water: 0.42.
Test vessel per concentration: 1.
Photoperiod 16hrs light, 8hrs darkness.
Test temperature: 12°C.
Feeding: None.

Reference substance (positive control):

not required

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)

Remarks on result:

other: Negative

Remarks:

0 mortalities

Details on results:

When the 10 fish were compared with the 10 control fish, there were no mortalities and no clinical symptoms observed at any concentration.

Reported statistics and error estimates:

No statistical analysis was carried out since no lethality was observed at the highest tested concentration. The median lethal concentration (LC50) was above this value.

Sublethal observations / clinical signs:

MORTALITY AND SYMPTOMS

 

Cumulative mortality (number of dead animals):

 

Nominal concentration (mg/L)	Number of fish	1 hour	4 hours	24 hours	48 hours	72 hours	96 hours
0	10	0	0	0	0	0	0
100	10	0	0	0	0	0	0

 

Symptoms:

 

Nominal concentration (mg/L)	Number of fish	1hour	4hours	24hours	48hours	72hours	96hours
0	10	n	n	n	n	n	n
100	10	n	n	n	n	n	n

Explanation of abreviations

n = no symptoms observed                           T = tottering                            

D = swimming at the bottom                         C Swimming at the surface

A = apathy

Numbers in brackets = number of animals affected

Effect concentrations (mg/L) based on nominal concentrations of the technical test substance:

 

	Observation time(hours after start of exposure)
	1hour	4hours	24hours	48hours	72hours	96hours
LCO	100	100	100	100	100	100
LC50 (95% confidence limits)	>100 -	>100 -	>100 -	>100 -	>100 -	>100 -
LC100	>100	>100	>100	>100	>100	>100
NOEC	100	100	100	100	100	100

Validity criteria fulfilled:

yes

Conclusions:

The 96 hour study showed no signs of toxicity (LC50) for glycoluril in the rainbow trout at the tested concentration (100 mg/L) it was therefore concluded the LC50 is above 100mg/L.

Executive summary:

The study aimed to assess the acute toxic effects of the test substance glycoluril in rainbow trout (Oncorhynchus mykiss) over a 96 hour exposure period. The study was conducted according to EPA-SEP 504, OECD Guideline 203 (1992) and EEC directive 92/69, Annex V, C1. 10 fishes were observed for survival and toxic signs within 1 hour after start exposure and 4,24,48,72, and 96 hours after start exposure to the test substance.

No lethality was observed at the highest tested concentration. The median lethal concentration (LC50) was above this value. And so, it was concluded that the 96 hour LC50 value for the test substance in the rainbow trout was > 100 mg/L based on analytically confirmed nominal concentrations. The substance does not meet the classifcation criteria for acute toxicity to fish under Regulation (EC) No. 1272/2008.

Description of key information

To determine the acute toxicity of the test substance in rainbow trout (Oncorhynchus mykiss) the following the guidelines were used; OECD 203 (July 1992), EEC Directive 92/69, Annex V, C1, EPA-SEP No. 540/9 -85 -006 (1985), EPA Guideline Paragraph 72 -1 (1982), the tests were carried out in a laboratory with a GLP Certificate. Twenty (20) fish were tested (10 as a control group while 10 were tested with the test substance at a concentration of 100 mg/L). They were assessed for 96 hours after given a 14 day acclimatisation period to find the LC50. Clinical symptoms were also assessed and recorded. At the end of the test, period no mortalities were recorded and the test animals showed no sign of distress or illness.

It was therefore concluded that the test substance is non-toxic to fresh water fish; LC50 > 100 mg/L based on analytically confirmed nominal concentrations and therefore does not meet the classification criteria under Regulation (EC) No. 1272/2008.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L