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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study with GLP.
Qualifier:
according to
Guideline:
EPA OPP 72-2 (Aquatic Invertebrate Acute Toxicity Test)
Deviations:
not specified
Qualifier:
according to
Guideline:
other: Committee on Methods for Toxicity Tests with Aquatic Organisms. Methods of acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians. 1975. U.S. EPA, Ecol. Res. Ser. 660/3-75-009. as well as American Public Health Association. 1980.
Deviations:
not specified
Qualifier:
equivalent or similar to
Guideline:
EU Method C.2 (Acute Toxicity for Daphnia)
Deviations:
not specified
GLP compliance:
yes
Analytical monitoring:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: dilution in Daphnid test water.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna
- Source: Cultured at the ABC facilities
- Age at study initiation: less than 24 h old
- Feeding during test: no

ACCLIMATION
- Acclimation period: no data
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
none.
Hardness:
CaCO3: 206-275 ppm
Test temperature:
19 - 20 °C
pH:
between 8.0 and 8.4
Dissolved oxygen:
between 8.1 and 8.7 mg/L
Salinity:
freshwater
Nominal and measured concentrations:
nominal concentrations: Control (0 mg/L), 56mg/L, 100mg/L, 180mg/L, 320mg/L, 560mg/L, and 1000 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: 250 mL glass beakers filled with 200 mL daphnid culture/test water, covered with loose-fitting petri dish covers.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source: Well water, used by ABC's Aquatic Toxicity Division
- Total organic carbon: 1.4 ppm
- Culture medium different from test medium: no


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours daylight with 30 minutes simulated down dawn and dusk periods.
- Light intensity: 50 - 70 footcandeles


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observation all 24 hours for mortality and abnormal effects such as surfacing, clumping of the daphnids together and daphnids tending to the bottom of test chambers.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: logarithmic series from 56 to 1000 mg/L
- Range finding study
- Test concentrations: 1.0, 10, 100, and 1000mg/L.
- Results used to determine the conditions for the definitive study: yes.
Reference substance (positive control):
not specified
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
24 h
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
200 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks:
and behaviour
Remarks on result:
other: 95% CL: 160 to 250 mg/L
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
400 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks:
and behaviour
Remarks on result:
other: 95% CL: 330 - 480 mg/L
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
< 56 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Remarks:
and behaviour
Details on results:
- Mortality of control: 0%
- Other adverse effects control: no
Results with reference substance (positive control):
n.a.
Reported statistics and error estimates:
The confidence limit EC50 24h: 330 -480 mg/L and EC50 48h: 160 - 250 mg/L

 Nominal Concentr. (mg/L)  Percent Mortality    Percent Mortality  Percent adversely affected      Percent adversely affected
   24 h  48 h  24 h  48 h
 Control  0  0  0  0
 56  0  0  0  10
 100  0  5  0  10
 180  0  20  0  50
 320  5  10  60  65
 560  0  5  60  95
 1000  5  20  95  100
Validity criteria fulfilled:
not specified
Conclusions:
The toxicity of melamine to Daphnia magna is low.
Executive summary:

The toxicity of the melamine to Daphnia magna was determined in a static test over 48h.

The 24 h and 48 h LC 50 values were both estimated to be > 1000 mg/L. The 24 and 48 hour EC 50 values were 400 and 200 mg/L respectively.

The no observed effect concentration (NOEC) was found to be <56 mg/l.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
07/03/1990-09/03/1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study is conducted to GLP and in compliance with agreed protocols, with no deviations from standard test guideline.
Qualifier:
according to
Guideline:
EPA OTS 797.1300 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)
Deviations:
yes
Remarks:
One concentration and a no-treatment control were tested in triplicate. a control exposure to determine the effects of pH on test organisms was not conducted. Ph and dissolved oxygen concentration were not measured at 24 hr.
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
not specified
Details on sampling:
- Concentrations: samples were collected from the control treatment groups at 0 and 48 hrs.
Vehicle:
yes
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances):
- Controls: vehicle
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): filtered (milli-Q filter) deionized, well water
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia magna (water flea)
- Source: Stock cultures maintained at ESE.
- Age at study initiation (mean and range, SD): <24hr
- Feeding during test - no

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
No
Hardness:
188 mg/L as CaCO3 at 0h
Test temperature:
20 ± 2°C
pH:
Control test:7.7-8.2 (measured at 0 and 48hr)
Test substance: 8.0-8.3 (measured at 0 and 48hr)
Dissolved oxygen:
Control test: 8.0-9.7 mg/L (measured at 0 and 48hr)
Test substance: 8.0-9.6 mg/L (measured at 0 and 48hr)
Salinity:
Not applicable
Nominal and measured concentrations:
Control:
nominal concentration: 0 mg/L

Test:
nominal concentration: 100 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: cylindical, glass, exposure chambers ( 100 mm diameter X 50 mm height)
- Type: closed
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 3
- No. of vessels per vehicle control (replicates): 3



TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Milli-Q water. Pumped from a deep well and passed through activated charcoal, cation, anion and mixed beds and then through the Milli-Q system prior to use. Sanple collected on 8 March 1990.
-pH: 7
- Total organic carbon: 1.3 mg/L
- Metals:
arsenic: <2.3 µg/L
aluminium: <27.5 µg/L
cadmium: <0.5 µg/L
calcium: 0.3 mg/L
chromium: <7.1 µg/L
cobalt: <16.4 µg/L
copper: <3.6 µg/L
iron: 23.3 µg/L
lead: < 1.4 µg/L
mercury:<0.2 µg/L
magnesium: 0.5 µg/L
nickel: <26.0 µg/L
selenium: <1.8 µg/L
silver: <6.8 µg/L
sodium: 3.0 µg/L
zinc: 8.3 µg/L
- Pesticides:
Pesitcides-Organochlorine & PCB's: <0.006 µg/L
Organophosphate: <0.088 µg/L

COD: 6.0 mg/L

filterable solids: 13.0 mg/L

OTHER TEST CONDITIONS
- Photoperiod: 16-hours light and 8-hours dark.
- Light intensity: no data


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Test organisms were observed daily for immobilisation (upon prodding) and any behavioural changes , and paralysed organisms removed.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: not applicable
- Justification for using less concentrations than requested by guideline: Results of range-finding study.
- Range finding study
- Test concentrations: up to 100 mg/L
- Results used to determine the conditions for the definitive study: In a range-finding test, mortalities occured down to 0.1 mg test substance/L of exposure water. However it was believed that mortalities occured due to the possible contamination of exposure chambers. Thererfore, in the definitive test, the test substance was used to provide nominal exposure concentrations of 0 mg/L (dilution water control) and 100 mg/L
Reference substance (positive control):
yes
Remarks:
sodium doceyl sulfate
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
based on test material purity of 100%
Basis for effect:
mobility
Remarks:
immobilisation
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
based on test material purity of 100%
Basis for effect:
mobility
Remarks:
immobilisation
Details on results:
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: A precipitate was noted in two treatment exposures at 48 hr.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
Statistical analysis (t-test) of data indicated there was no significant (alpha + 0.05) difference between the number of daphnids immobilised in control and treatment exposures.

 

Table 1: Immobility of water fleas (D. magna) exposed to tetrapotassium pyrophosphate in a 48-hr static , acute toxicity test.

 

Nominal concentration of tetrapotassium pyrophosphatea

(mg/L)

Cumulative number of immobilised organisms and

Percent mortality (parenthic value)

24 hr

48 hr

0 (control)

0(0)

0(0)

100

0(0)

2(7)

 

a Cacluated based on 100% puroty of the test substance. Source: ESE 1990.

Validity criteria fulfilled:
yes
Conclusions:
The test substance, tetrapotassium pyrophosphate, was not acutely toxic to the water flea, Daphnia magna, at a nominal concentration of 100 mg/L.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Remarks:
Summary of available data used for the endpoint assessment of the target substance
Adequacy of study:
weight of evidence
Justification for type of information:
Please refer to the analogue justification attached to section 13
Reason / purpose:
read-across source
Reason / purpose:
read-across source
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: Source, WoE, RA-A, CAS 108-78-1, Frazier 1988
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Remarks:
based on test material purity of 100%
Basis for effect:
mobility
Remarks on result:
other: Source, WoE, RA-A, CAS 7320-34-5, Vaishnav 1991

Description of key information

No toxic effects on aquatic invertebrates were recorded within the limit of water solubility of the substance.

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
100 mg/L

Additional information

Data on the acute toxicity of the target substance diphosphoric acid, compound with 1,3,5-triamine (1:2) to fish are not available. The evaluation of the acute toxicity is based on a weight of evidence approach conducted with available data on the source substances melamine (CAS 108-78-7) and tetrapotassium pyrophosphate (CAS 7320-34-5). Whereby melamine represents the 1,3,5-triamine moiety of the target substance. The source substance tetrapotassium pyrophosphate was selected to assess the toxicity of the diphosphoric acid moiety. The read across approach is in accordance with Regulation (EC) No. 1907/2006, Annex XI, 1.5. Further justification is given in the analogue justification in IUCLID section 13.

Comparable results were obtained in the two available studies on the source substance melamine (CAS 108-78-7). In a guideline study following EPA OPP 72-2 Daphnia magna was exposed to nominal concentrations of 56, 100, 180, 320, 560 and 1000 (Frazier, 1988). The determined LC50 (48 h) was > 1000 mg/L, the reported EC50 (48 h) was 200 mg/L.

The acute toxicity of tetrapotassium pyrophosphate to aquatic invertebrates was investigated in a study following EPA OTS 797.1300 (Vaishnav et al., 1991). The study was conducted as a limit test under static conditions. Daphnia magna was exposed to a nominal test substance concentration of 100 mg/L for 48 hours. At test termination an EC50 (48 h) >100 mg/L (nominal) was determined.

Based on the available data for the source substances the target substance diphosphoric acid, compound with 1,3,5-triamine (1:2) is considered not acutely toxic to aquatic invertebrates.