1H-1,4,7-Triazonine, octahydro-1,4,7-trimethyl-

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 February 1993 to 18 February 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted on read-across material

Justification for type of information:

The read-across justification will be provided as soon as possible.

Reason / purpose for cross-reference:

other: Read-across target

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- The test solutions were analysed by Analytical Chemistry.
- Sample from Vessels 1 and 3 (0 mg/L) and 5 and 7 (97.5 mg/L) were measured at 0, 24 (new and old) and 48 hours.
- The analysis was conducted according to the method described in MSD Analytical Chemistry SOP 404 01 and when possible was carried out on the day of sampling. If this was not possible the samples were stored at 4 °C until analysed.

Vehicle:

no

Details on test solutions:

A 975 mg/L stock (taken as 1000 mg/L during the study) was prepared by adding 0.25 g of the test material to 250 mL distilled water. Test solutions were prepared by diluting the stock with ASTM medium-hard medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Cultures were established with 10 <24 hours old neonates in 800 mL of medium; when these Daphnia become gravid their numbers are reduced to 5 individuals. Cultures are reset with <24 hours old neonates when the adults are 24 days old.

ACCLIMATION
- Cultures were maintained in a synthetic medium (ASTM medium-hard medium) with the addition of an organic seaweed-derived extract. ASTM has the same chemical composition and similar pH and dissolved oxygen concentration as that used to prepare test solutions.
- Daphnia cultures were fed cells of the green alga ChIorella vulgaris.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

164 mg/L (as CaCO3)

Test temperature:

20 ± 2 °C

pH:

7.81

Dissolved oxygen:

8.78 mg/L

Nominal and measured concentrations:

Nominal: 97.5 mg/L
Mean Measured: 93.5 (0 hours), 101.7 (24 hours new), 100.5 (24 hours old) and 102.1 (48 hours) mg/L.

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates):4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ASTM medium-hard medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: The temperature, pH, dissolved oxygen concentration and total hardness (as mg/L CaCO3) of selected test solutions were measured at 0, 24 and 48 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hour light, 8 hour dark photoperiod
- Light intensity: normal laboratory lighting

EFFECT PARAMETERS MEASURED:
- Immobility was recorded at 24 and 48 hours

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 97.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

TEST CONDITIONS
- The results of the analysis of test conditions and synthetic medium indicate that temperature, pH, dissolved oxygen and total hardness were maintained within their standard ranges and therefore the test was qualitatively acceptable.

TEST CONCENTRATIONS
- No test material was found in the control group samples.
- In the test material group (nominal 97.5 mg/L) mean measured concentrations were: 93.5 (0 hours), 101.7 (24 hours new), 100.5 (24 hours old) and 102.1 (48 hours) mg/L. The test material concentrations were therefore maintained during the test.

IMMOBILITY
- At 24 and 48 hours, some of the daphnia exposed to the test material were immobilised in the surface film. No other immobile daphnia were observed in these
vessels, suggesting that immobility may not have been a direct response to the inherent toxicity of the test material.

Table 1: Cumulative Percentage Immobile at 24 and 48 Hours

Nominal Concentration (mg/L)	24 Hours	48 Hours
0	0	5
97.5	0	30

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of this study the 48 hour EC50 of the test material to Daphnia magna was greater than 97.5 mg/L.

Executive summary:

The acute toxicity of the test material to aquatic toxicity was investigated in accordance with the standardised guidelines OECD 202 and EU Method C.2., under GLP conditions.

The test was conducted under static renewal conditions. Daphnia were exposed to one concentration of 97.5 mg/L. Analysis of the test material indicated that the exposure concentration remained stable throughout the test.

There was a 30 % mortality at 48 hours. These Daphnia appeared to have died as a result of being caught in the surface film rather than as a direct response to the inherent toxicity of the test material.

Under the conditions of this study the 48 hour EC50 of the test material to Daphnia magna was greater than 97.5 mg/L.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 October 1993 to 20 October 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study conducted on read-across material

Justification for type of information:

The read-across justification will be provided as soon as possible.

Reason / purpose for cross-reference:

other: Read across target

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Samples of test solution were taken at 0, 24 (old and new) and 48 hours for determination of test material concentrations. The test solutions were analysed by Analytical Chemistry. The analysis was carried out according to MSD SOP 404.

Vehicle:

no

Details on test solutions:

The stock solution was prepared by weighing out 2.05 g of test material and dissolving in 2.0 litres ASTM "medium-hard" medium making a 1.0 g/L stock. Test
solutions were prepared by adding known volumes of stock to ASTM using an electronic pipette.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Cultures were reset using 10 neonates, when they become gravid their numbers were reduced from 10 to 5 and the production of neonates results in their daily removal.

ACCLIMATION
- Cultures were maintained in a synthetic medium with the same chemical composition and similar pH and dissolved oxygen concentration as that used to prepare test solutions. Culture medium consists of 800 mL ASTM medium-hard medium with the addition of an organic additive, derived from the marine macroalga Ascophyllum nodosum.
- The cultures were fed daily with cells of the green algae ChIarella vulgaris.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

171 mg/L (as CaCO3)

Test temperature:

17 - 20 °C

pH:

8.0

Dissolved oxygen:

8.6 mg/L

Nominal and measured concentrations:

Nominal: 32, 56, 100, 180 and 320 mg/L
Measured (0 hours): 34.2, 57.5, 99.8, 178.4 and 331.8 mg/L
Measured (24 hours, new): 33.3, 57.4, 100.6, 158.4 and 278.5 mg/L
Measured (24 hours, old): 32.9, 52.0, 98.5, 174.4 and 325.9 mg/L
Measured (48 hours): 31.8, 56.2, 99.9, 175.4 and 318.3 mg/L

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates):4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ASTM medium-hard medium
- Culture medium different from test medium: no
- Intervals of water quality measurement: The temperature, pH, dissolved oxygen concentration and total hardness (as mg/L CaCO3) of selected test solutions were measured at 0, 24 and 48 hours.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hour light, 8 hour dark photoperiod
- Light intensity: normal laboratory lighting

EFFECT PARAMETERS MEASURED:
- Immobility was recorded at 24 and 48 hours

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 320 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

56 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 320 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

TEST CONDITIONS
- The results of the analysis of test conditions and synthetic medium indicate that temperature, pH, dissolved oxygen and total hardness were maintained within their standard ranges and therefore the test was qualitatively acceptable.

TEST CONCENTRATIONS
- Mean concentrations were between 95 and 103 % of nominal values. Effect concentrations are, therefore, based on nominal concentrations.

IMMOBILITY
- Results can be seen in Table 1.
- At all test concentrations except the control some Daphnia were caught in the surface film.

Table 1: Cumulative Percentage Immobile at 24 and 48 Hours

Nominal Concentration (mg/L)	24 Hours	48 Hours
0	0	55
32.0	0	20
56.0	0	20
100.0	5	10
180.0	5	25
320.0	9	27.3

 

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of this study the 24 and 48 hour EC50 of the test material to Daphnia magna was greater than 320 mg/L. The 24 hour NOEC was 56.0 mg/L.

Executive summary:

The acute toxicity of the test material to aquatic toxicity was investigated in accordance with the standardised guidelines OECD 202 and EU Method C.2., under GLP conditions.

The test was conducted under static renewal conditions. Daphnia were exposed to a range of concentrations from 32.0 – 320 mg/L. Analysis of the test material indicated that the exposure concentration remained stable throughout the test. Mean concentrations were between 95 and 103 % of nominal values. Effect concentrations are, therefore, based on nominal concentrations.

At all test concentrations except the control some Daphnia were caught in the surface film.

Under the conditions of this study the 24 and 48 hour EC50 of the test material to Daphnia magna was greater than 320 mg/L. The 24 hour NOEC was 56.0 mg/L.

Description of key information

Marshall (1993) AT/501/03

Under the conditions of this study the 48 hour EC50 of the test material to Daphnia magna was greater than 97.5 mg/L.

Marshall (1993) AT/501/06

Under the conditions of this study the 24 and 48 hour EC50 of the test material to Daphnia magna was greater than 320 mg/L. The 24 hour NOEC was 56.0 mg/L.

Additional information

Marshall (1993) AT/501/03

The acute toxicity of the test material to aquatic toxicity was investigated in accordance with the standardised guidelines OECD 202 and EU Method C.2., under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

The test was conducted under static renewal conditions. Daphnia were exposed to one concentration of 97.5 mg/L. Analysis of the test material indicated that the exposure concentration remained stable throughout the test.

There was a 30 % mortality at 48 hours. These Daphnia appeared to have died as a result of being caught in the surface film rather than as a direct response to the inherent toxicity of the test material.

Under the conditions of this study the 48 hour EC50 of the test material to Daphnia magna was greater than 97.5 mg/L.

Marshall (1993) AT/501/06

The acute toxicity of the test material to aquatic toxicity was investigated in accordance with the standardised guidelines OECD 202 and EU Method C.2., under GLP conditions. The study was awarded a reliability score of 1 in accordance with the criteria set forth by Klimisch et al. (1997).

The test was conducted under static renewal conditions. Daphnia were exposed to a range of concentrations from 32.0 – 320 mg/L. Analysis of the test material indicated that the exposure concentration remained stable throughout the test. Mean concentrations were between 95 and 103 % of nominal values. Effect concentrations are, therefore, based on nominal concentrations.

At all test concentrations except the control some Daphnia were caught in the surface film.

Under the conditions of this study the 24 and 48 hour EC50 of the test material to Daphnia magna was greater than 320 mg/L. The 24 hour NOEC was 56.0 mg/L.