N-cyclopropyl-1,3,5-triazine-2,4,6-triamine

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:

2003/07/21 to 2003/09/04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Study performed under GLP conditions. The guidelines followed in this study are OECD guideline 202 part 1 91884, JMAFF (2-7-2-1) (2001) and EU Commission Directive 92/69/EEC C.2 (1992). No deviations occurred from these guidelines.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

Part 1 91884

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: JMAFF

Version / remarks:

2-7-2-1, 2001

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N-cyclopropyl-1,3,5-triazine-2,4,6-triamine

EC Number:

266-257-8

EC Name:

N-cyclopropyl-1,3,5-triazine-2,4,6-triamine

Cas Number:

66215-27-8

Molecular formula:

C6H10N6

IUPAC Name:

N2-cyclopropyl-1,3,5-triazine-2,4,6-triamine

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

The test material is Cyromazine, it is a white, light beige solid crystalline powder.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Five Daphnia magna (6-24 hours old and not first progeny brood) were added to each of four replicate vessels for each treatment and control. The daphnids were not fed for the duration of the test. The Daphnia magna were assessed visually for signs of immobility after 24 and 48 hours. All samples were deep-frozen (at about -20 °C) immediately after.

Test solutions

Vehicle:

yes

Remarks:

reconstituted water

Details on test solutions:

The test medium of the highest nominal concentration of 100 mg/L was prepared by dissolving 100 mg of test item completely in 1000 mL of test water using ultrasonic treatment for 15 minutes and intense stirring for 15 minutes at room temperature. In a series of different dilution steps this test medium was diluted with test water to prepare the test media with the lower test item concentrations as stated above. The test media were prepared just before introduction of the Daphnia magna (= start of the test).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

- Age at study initiation: 6-24 hours old and not first progeny brood
- Diet: Not fed during test

Study design

Test type:

static

Water media type:

other: reconstituted water

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

2.5 mmol/L as CaCO3

Test temperature:

water was at constant 20°C

pH:

7.6 to 7.7

Dissolved oxygen:

8.5 mg/L

Nominal and measured concentrations:

Nominal Concentrations: 1.0, 2.2, 4.6, 10, 22, 46, 100 mg/L

Details on test conditions:

Test vessel consisted of a 100mL glass beaker (with a lid) containing 50 mL of the appropriate test solution. The water was aerated until oxygen saturation prior to initiation but for the 48-hours duration of the test the vessels were not aerated. Analytical determination of the test substance concentrations were made at test start and end.

Reference substance (positive control):

yes

Remarks:

Control in this experiment was untreated reconstituted water

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

46 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 4

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

The measured concentrations at test start or at test end were 99% to 108% of nominal All results are therefore presented in terms of nominal concentration
Immobilisation and symptoms of toxicity

Any other information on results incl. tables

Physical parameters monitored:

The water was at a constant 20°C and measured light intensity ranged from 460-670 lux (16-hour photoperiod).  The pH of the test medium and the control ranged from 7.6 to 7.7 and the water hardness was 2.5 mmol/L as CaCO₃. Dissolved oxygen concentration in the test media was at least 8.5 mg/L.

 

Analytical findings:

The measured concentrations at test start or at test end were 99% to 108% of nominal All results are therefore presented in terms of nominal concentration

 

Immobilisation and symptoms of toxicity:

 

Table 1

Acute toxicity of test material on the mobility of Daphnia magna in a 48-hour static study

 

Nominal concentration (mg/L)	Immobilised Daphnia magna after 24 hours		Immobilised Daphnia magna after 48 hours
	No.	%	No.	%
Control	0	0	0	0
1.0	0	0	0	0
2.2	0	0	0	0
4.6	0	0	0	0
10	0	0	4	20
22	0	0	5	25
46	0	0	7	35
100	0	0	7	35

 

The 48-h NOEC was determined to be 46 mg cyromazine/L and the 48-h LOEC was 10 mg cyromazine/L.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48-hour EC50 of cyromazine with Daphnia magna was >100 mg/L.

Executive summary:

The acute toxicity of the test item cyromazine to Daphnia magna was determined in a 48-hour static test according to the JMAFF Test Guideline, 2-7-2-1 (2001), the EU Commission Directive 92/69/EEC, Part (1992), and the OECD Guideline for Testing of Chemicals, No. 202, Part I (1984). This study followed the procedures indicated by the following internationally accepted guidelines and recommendations.

The nominal test item concentrations tested were 1.0, 2.2, 4.6, 10, 22, 46, and 100 mg/L. Additionally, a control was tested in parallel. During the test period the test item concentrations in the test media were measured in the range from 99 to 108 % of the nominal values. Under the conditions of the test the test item

Cyromazine was stable in the test media during the test period of 48 hours. The 24-hour EC50 was > 100 mg/L along with the 24 hour EC100. The 48 hour EC50 was > 100 mg/L Therefore, all reported results are related to the nominal concentrations of the test item.