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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:
key study
Study period:
10 February to 12 February 1979
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Non-GLP, not conducted according to recognised guideline, but fully detailed method and results.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Hardness and alkalinity were ascertained by standard analytical prodedures (American Public Health Association, 1976). Test practices followed those recommended by the Committee on Mebhods for Toxicity Tests with Aquatic Organisms (1975).

Test practices followed those recommended by the Committee on Methods for Toxicity Tests with Aquatic Organisms (1975). A working stock solution of the test material in reagent grade acetone, 100 mg/ml, was prepared by weight to a precision of 0.1 mg. Secondary stocks (1, 10 mg/ml) were prepared by serial dilution. Six concentrations, a control and solvent control were used, with four replicates of each. Test vessels were 250 ml glass beakers. Testing was begun by thoroughly mixing measured volumes of stock solution and dilution water in one-liter volumetric flasks, to yield nominal test material concentrations of 1.0, 1.8, 3.2, 5.6, 10.0 and 18.0 mg/l.mg/l. Two hundred ml of each concentration was decanted into each of four test beakers; four additional beakers, as controls, contained 200 ml each of 100% dilution water, and four others, as solvent controls, each contained 200 ml solution of acetone in dilution water, at the same acetone level as in the highest test concentration.

Five.organisms were randomly introduced into each of the 24 test, 4 control and 4 solvent control beakers. The tray of beakers was held for the duration of the test in a refrigerator incubator at a constant temperature of 20 + 1°C. Dissolved oxygen and pH were determined initially and at 48 hours for all test concentrations and the controls. Mortalities were recorded at 24 and 48 hours .

The concentration of test material lethal to 50% of the population (LC50, the Median Lethal Concentration) and 95% confidence limits were determined for the 48 hour exposure period by the Spearman - Karber Estimator (Finney, 1971), using mortality data from 1.0 mg/l through 10.0 mg/l treatments.

References to books used:
- American Public Health Association. 1976. Standard Methods for the
Examination of Water and Wastewater
- Committee on Methods forToxicity Tests with Aquatic Organisms. 1975. Methods for Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians.
- Finney, D. J. 197 1. Statistical Methods in Biological Assay.
GLP compliance:
not specified
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable
Analytical monitoring:
no
Details on sampling:
Not applicable
Vehicle:
not specified
Details on test solutions:
A working stock solution of the test material in reagent grade acetone, 100 mg/ml, was prepared by weight to a precision of 0.1 mg. Secondary stocks (1, 10 mg/ml) were prepared by serial dilution. Six concentrations, a control and solvent control were used, with four replicates of each. Test vessels were 250 ml glass beakers. Testing was begun by thoroughly mixing measured volumes of stock solution and dilution water in one-liter volumetric flasks, to yield nominal test material concentrations of 1.0, 1.8, 3.2, 5.6, 10.0 and 18.0 mg/l.mg/l. Two hundred ml of each concentration was decanted into each of four test beakers; four additional beakers, as controls, contained 200 ml each of 100% dilution water, and four others, as solvent controls, each contained 200 ml solution of acetone in dilution water, at the same acetone level as in the highest test concentration.
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water Flea
- Strain: Daphnia Magna Straus
- Source: A UCES laboratory stock culture, the original population having been obtained from the National Water Quality Laboratory in Duluth, Minnesota.
- Age at study initiation (mean and range, SD): No data.
- Weight at study initiation: No data.
- Length at study initiation (length definition, mean, range and SD): No data.
- Valve height at study initiation, for shell deposition study (mean and range, SD): No data.
- Peripheral shell growth removed prior to test initiation: No data.
- Feeding during test: One hour before the test they were fed, and no additional food was administered thereafter.
- Food type: No data.
- Amount: No data.
- Frequency: No data.

ACCLIMATION
- Acclimation period: No data.
- Acclimation conditions: No data.
- Type and amount of food: No data.
- Feeding frequency: No data.
- Health during acclimation (any mortality observed): No data.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Post exposure observation period:
All observations were made during the exposure period.

Hardness:
232 mg/l as CaCO3
Test temperature:
20 +/- 1 deg C
pH:
7.51
Dissolved oxygen:
The concentration of dissolved oxygen was measured throughout but no mean or required value was given.
Salinity:
Not applicable.
Conductivity:
Not specified
Nominal and measured concentrations:
Six concentrations, a control and solvent control were used. Nominal concentrations were: 1.0, 1.8, 3.2, 5.6, 10.0 and 18.0 mg/l.

Details on test conditions:
TEST SYSTEM
- Test vessel: Beaker
- Type (delete if not applicable): open / closed
- Material, fill volume: Glass, 250ml.
- Aeration:. The dilution water was stored in a 95 liter glass reservoir and was vigorously aerated before use.
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 5, randomly selected.
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: no data.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The dilution water used in the test was obtained from the well on the Tarrytown site of UCES, since first instar daphnids readily survive in water from the same source for 48 hours without feeding.
- Alkalinity: 147 mg/ l as CaCO3
- Hardness: 232 mg/l as CaCO3
- Conductance: 600 umhos/cm

OTHER TEST CONDITIONS
- Adjustment of pH: no data.
- Photoperiod: no data.
- Light intensity: no data.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Dissolved oxygen and pH were determined initially and at 48 hours for all test concentrations and the controls.
- Mortalities were recorded at 24 and 48 hours .

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Various.
- Range finding study: Yes- a preliminary test was carried out.
- Test concentrations: 1.0, 1.8, 3.2, 5.6, 10.0 and 18.0 mg/l.
- Results used to determine the conditions for the definitive study: No data.
Reference substance (positive control):
yes
Remarks:
Sodium lauryl sulfate (SLS)
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
1.5 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: 95% CL
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
The LC50 values for the registered substance after 24 and 48 hours are 5.97 and 1.50 mg/L respectively. The NOEC value was determined to be 1.00mg/L.
No other data was detailed within the report.
Results with reference substance (positive control):
- Results with reference substance valid?
- Mortality: Yes
- LC50: 3.36 mg/L.

Results:

The 48 hour LC50 with 95% confidence limits for K-200 to Daphnia magna is 1.50 mg/L (1.34 - 1.67 mg/L). The 48 hour no observed to be 1.00 mg/l. It should be noted that LC50 values may vary with different species, temperatures and water qualities.

Table 1: Percentage Mortalities:

Control 

Solvent Control 

Test 

Material 

Nominal 

Concen 

tration 

mg/L 

 

 

 

1.00 

1.80 

3.20 

5.60 

10.0 

18.0 

24 hour 

5

20 

45

70

95 

48 hour 

85

95

100

100

100 

Table 2: LC50 Values:

 

24 hour 

48 hour 

LC50 

mg/L 

5.97

1.50

95% Confidence 

Low 

4.82

1.34

Limits 

High 

7.41

1.67 

Table 3: Dissolved Oxygen (mg/L):

Control 

Solvent Control 

Test 

Material 

Nominal 

Concen 

tration 

mg/L 

 

 

 

1.00

1.80 

3.20 

5.60

10.0 

18.0 

Initial 

10.0

9.8

10.0

10.2

10.2

10.0

10.2

10.2 

48 hour 

9.4

9.0

9.2 

9.0

9.0 

9.0 

8.8 

8.8

Table 4: pH:

Control 

Solvent Control 

Test 

Material 

Nominal 

Concen 

tration 

mg/L 

 

 

 

1.00

1.80 

3.20 

5.60 

10.0 

18.0 

Initial

7.51

7.74

7.70 

7.75

7.71 

7.74 

7.71 

7.75 

48 hour

 8.31

8.37 

8.38 

8.39

8.43 

8.49 

8.49 

8.53

Validity criteria fulfilled:
not specified
Conclusions:
This test was conducted on a sample of the material where the content of Triphenyl phosphate is present at > 5%

The LC50 values for the registered substance after 24 and 48 hours are 5.97 and 1.50 mg/L respectively. The NOEC value was determined to be 1.00mg/L.
Executive summary:

This test was conducted on a sample of the material where the content of Triphenyl phosphate is present at > 5%

The short-term toxicity of the registered substance to Water Flea (Daphnia Magna Straus) was assessed, with the 96-hour LC50 value determined to be 1.50 mg/L and the NOEC to be 1.00 mg/L.

On the basis of the study results, the substance should be classified as "toxic" to aquatic organisms.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
31 July 2000 - 07 April 2001
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Study, conducted to recognised guidelines.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable.
Analytical monitoring:
yes
Details on sampling:
For the purpose of chemical analysis during the limit test, single aliquots (ca 50 ml) of centrifuged solution were removed from test and control solutions and taken for analysis at 0 h and 24 h as fresh test solution. At 24 h and 48 h an aliquot (ca 15 ml) was removed from each replicate test vessel, including controls, pooled at each concentration and taken for analysis as expired test solution.

Test samples were analysed according to the procedure established and validated under Inveresk Protocol No. 281518.
Vehicle:
no
Details on test solutions:
A preliminary range finding test was conducted over a 48 h period under semi-static conditions, using the following nominal loading rates of Durad 310M in Daphnia medium, prepared as water accommodated fraction's (WAF's): 0, 1, 10, 100 and 1000 mg/l.

Individual WAF's were prepared by adding weighed amounts of Durad 310M (0.97-1.15, 9.96-9.99, 99.85-99.87 and 999.91 mg) to Daphnia medium (1 litre) and stirring this in the dark for an optimal period of 1 h. In the stability trial (conducted under Inveresk Project No. 281518) a period of 1 h stirring was shown to result in the maximum achievable concentration of test item in test medium and the lowest level of degradation. Based on preliminary investigations and observations in the stability trial, WAF's were prepared in 1 litre volumetric flasks and not 2 litre containers as stated in the protocol. This protocol deviation does not affect the validity of the study. The rate of stirring was adjusted to ensure the vortex was <5% of the static depth of mixture. Following the period of stirring, the flask contents were allowed to settle for 1 h after which ca 350 ml of solution was siphoned from the centre of the vessel, with the first ca 100 ml being discarded. The remaining ca 250 ml of solution was centrifuged (3000 rpm for 15 mins) and the supernatant used as the test solution. The test solutions were prepared as WAF then centrifuged so that the Daphnia were exposed only to the water soluble fraction.
Test organisms (species):
Daphnia magna
Details on test organisms:
Test Species
Daphnia magna (water flea) were used for this study. They were bred within the laboratory by acyclical parthenogenesis and the individuals used were between 6 and 24 h old. Daphnia cultures were maintained in a synthetic medium (Elendt M7) and all testing was done using this medium.

Food and Feeding Regime
Daphnia cultures were fed on a diet of axenic cultures of Pseudokirchneriella subcapitata (Strain 278/4, CCAP, Ambleside, Cumbria). Daphnia were not fed during the test.
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
48 h
Remarks on exposure duration:
Performed on WAF
Post exposure observation period:
None
Hardness:
196 mg/l as CaCO3
Test temperature:
The temperature ranged from 20.3-20.8 °C and did not deviate by more than 0.5 °C throughout the test period
pH:
pH ranged from 7.38-8.01
Dissolved oxygen:
Dissolved oxygen remained at or above 77% of air saturation value
Salinity:
Not applicable
Conductivity:
Not specified
Nominal and measured concentrations:
0, 1, 10, 100 and 1000 mg/l (nominal as WAF)
Details on test conditions:
Test Vessels
Glass crystallising dishes of ca 200 ml capacity and covered with perspex lids to prevent dust contamination and evaporation loss, were used as the test vessels.

Range Finding Test
A preliminary range finding test was conducted over a 48 h period under semi-static conditions, using the following nominal loading rates of Durad 310M in Daphnia medium, prepared as water accommodated fraction's (WAF's): 0, 1, 10, 100 and 1000 mg/l.

Individual WAF's were prepared by adding weighed amounts of Durad 310M (0.97-1.15, 9.96-9.99, 99.85-99.87 and 999.91 mg) to Daphnia medium (1 litre) and stirring this in the dark for an optimal period of 1 h. In the stability trial (conducted under Inveresk Project No. 281518) a period of 1 h stirring was shown to result in the maximum achievable concentration of test item in test medium and the lowest level of degradation. Based on preliminary investigations and observations in the stability trial, WAF's were prepared in 1 litre volumetric flasks and not 2 litre containers as stated in the protocol. This protocol deviation does not affect the validity of the study. The rate of stirring was adjusted to ensure the vortex was <5% of the static depth of mixture. Following the period of stirring, the flask contents were allowed to settle for 1 h after which ca 350 ml of solution was siphoned from the centre of the vessel, with the first ca 100 ml being discarded. The remaining ca 250 ml of solution was centrifuged (3000 rpm for 15 mins) and the supernatant used as the test solution. The test solutions were prepared as WAF then centrifuged so that the Daphnia were exposed only to the water soluble fraction.

Duplicate test vessels were prepared at each concentration, each containing 100 ml of test solution. Five Daphnia were added to each vessel within 30 min of preparation. Daphnia were transferred (by means of a wide bore glass pipette) to freshly prepared test solutions at 24 h.

Limit Test
Based on the results of the range finding test, the definitive test was conducted as a limit test. The limit test was conducted at the nominal loading rate of 1000 mg.l"1, with test solution prepared as water accommodated fraction (WAF). The test solution was centrifuged so that the Daphnia were exposed only to the water soluble fraction.
The WAF was prepared by adding a weighed amount of Durad 310M (0.99983-0.99990 g) to Daphnia medium (1 litre) and stirring this in the dark for an optimal period of 1 h. The rate of stirring was adjusted to ensure the vortex was <5% of the static depth of the mixture. Following the period of stirring, the flask contents were allowed to settle for 1 h after which ca 800 ml of solution was siphoned from the centre of the vessel, with the first ca 100 ml being discarded. The remaining ca 700 ml of solution was centrifuged (2500 rpm for 15 mins) and the supernatant used as the test solution.
Four test vessels were prepared at 1000 mg.l"1 with four control vessels included, each vessel containing 100 ml of test solution. Five Daphnia were added to each vessel within 30 min of preparation. Daphnia were transferred (by means of a wide bore glass pipette) to freshly prepared test solutions at 24 h.

Determination of Durad 310M in Daphnia Medium
For the purpose of chemical analysis during the limit test, single aliquots (ca 50 ml) of centrifuged solution were removed from test and control solutions and taken for analysis at 0 h and 24 h as fresh test solution. At 24 h and 48 h an aliquot (ca 15 ml) was removed from each replicate test vessel, including controls, pooled at each concentration and taken for analysis as expired test solution.
Test samples were analysed according to the procedure established and validated under Inveresk Protocol No. 281518.

Observations
The number of immobile Daphnia were recorded at 24 h and 48 h following the commencement of the tests. An animal was considered to be immobile if no movement was observed within 15 s following gentle agitation of the test vessel.

Test Solution Quality Parameters
Temperature, pH, conductivity and dissolved oxygen concentration were measured at 0, 24 and 48 h in each vessel during the limit test.
Dissolved oxygen and temperature were measured using a Mettler Toledo M0128 dissolved oxygen meter with dissolved oxygen and temperature probes, conductivity was measured with a Jencon's 4070 and a Mettler Toledo MC126 conductivity meter and pH with a Mettler Toledo MP120 pH meter.
Total hardness of the Daphnia medium was also measured and was determined to be 196 mg /l as CaCO3.

Environmental Control
Test vessels were maintained within the laboratory at a temperature of 18-22 °C. The temperature ranged from 20.3-20.8 °C and did not deviate by more than 0.5 °C throughout the test period. A light cycle of 16 h light and 8 h dark was in operation throughout the tests. Illumination was provided by artificial daylight fluorescent tubes.
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: immobilisation
Remarks on result:
other: Based on initial loading rates.
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: immobilisation
Remarks on result:
other: The No Observed Effect Loading (NOEL) for immobilisation was determined to be 1000 mg Durad 310M per litre
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
< 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
dissolved
Basis for effect:
other: observed effects
Remarks on result:
other: (<0.773 mg/l mean measured concentration).
Details on results:
Range Finding Test
The range finding test was conducted over a 48 h period under semi-static conditions, using the following nominal loading rates of Durad 310M in Daphnia medium, prepared as water accommodated fraction's (WAF's): 0, 1, 10,100 and 1000 mg/l.

After 24 h exposure, there was one (10%) immobile Daphnia recorded at 100 mg/l and one at 1000 mg/l. No other immobile Daphnia were recorded during the test.

Limit Test
Based on the results of the range finding test, the definitive test was conducted as a limit test. The limit test was conducted over a 48 h period under semi-static conditions at the nominal loading rate of 1000 mg Durad 310M / l Daphnia medium.
Test solutions were prepared as WAF then centrifuged so that the Daphnia were exposed to the water soluble fraction. All test solutions were clear and colourless throughout the test period.

Analytical samples were taken at 0 h and 24 h (fresh test solution) and 24 h and 48 h (expired test solution) to confirm the concentration of test material in solution (Table 2). The analytical results indicate that Durad 310M is only partially soluble in Daphnia medium under test conditions with fresh test solution concentrations ranging from 0.710-1.274 mg/l. Measured concentrations of expired test solution ranged from 0.436-0.671 mg/l indicating a loss of Durad 310M from the test medium under test conditions. Durad 310M was detected (limit of quantification=0.404 p.p.m.) in control samples at 0 h and 24 h (fresh test solution). The reason for this is unknown but may be an artefact of the system. Durad 310M was not detected in control samples at 24 h and 48 h (expired test solution) implying that control solutions were not contaminated with Durad 310M.

One immobile Daphnia was recorded at 1000 mg/l after 48 h exposure. This immobilisation is not regarded as significant. No other immobile Daphnia were recorded during the test (Table 3). It was noted at 24 h that Daphnia exposed to Durad 310M had material adhering to their bodies which impeded normal movement. This material is thought to be Durad 310M. By 48 h an increased amount of material had adhered to Daphnia exposed to Durad 310M and the one immobile Daphnia recorded was encased with this material. This material was not observed in the water or the test vessels and was not observed in the controls throughout the test. The results indicate that Durad 310M is not acutely toxic to Daphnia, under the conditions of the test. It is concluded that the EL50 of Durad 310M to Daphnia is >1000 mg/l. The No Observed Effect Loading (NOEL) for immobilisation is concluded to be 1000 mg Durad 310M / l. The NOEL for observed effects is <1000 mg/l (<0.773 mg/l mean measured concentration) as test item was observed to adhere to Daphnia and impede swimming ability.

All test solution quality parameters remained within acceptable limits throughout the test. The pH ranged from 7.38-8.01, conductivity from 0.54-0.58 mS and dissolved oxygen remained at or above 77% of air saturation value.
Results with reference substance (positive control):
Not applicable.
Reported statistics and error estimates:
None

Table 1

 

Durad 310M Determination of Acute Toxicity (EC50) to Daphnia (48 h, Semi-Static)

Cumulative Number of Immobile Daphnia Recorded During the Range Finding Test

 

Time

Tank Replicate

Nominal Loading of Durad 310M mg/l

 

 

(h)

 

0

1

10

100

1000

 

I

0

0

0

1

0

24

II

0

0

0

0

1

 

Per cent ImmobileDaphnia

0

0

0

10

10

 

I

0

0

0

1

0

48

II

0

0

0

0

1

 

Per cent immobileDaphnia

0

0

0

10

10

 

Five Daphnia at each of 2 replicates were tested at each concentration.

  

Table 2

Durad 310M Determination of Acute Toxicity (EC50) to Daphnia (48 h, Semi-Static)     Measured Concentrations of Durad 31 OM in Test Solutions During the Limit Test

 

Nominal Loading of Durad 310M (mg.r1)

Time (h)

Measured Concentration of Durad 310M (mg/l)

Mean Measured Concentration of Durad 310M (mg/l)

0

0

<LOQ

<LOQ/ND*

*24

ND

24

<LOQ

48

ND

 

 

 

 

1000

0

1.274

0.773

*24

0.671

24

0.710

*48

0.436

 LOQ = limit of quantification (limit of reliable quantification taken as lowest analytical standard = 0.404 ppm).

 * = Expired test solution ND = Not detected

The reason for measured concentrations of Durad 310M appearing in control samples is not known. It may be as a result of an artefact of the system. Durad 310M was not detected at 24 h and 48 h (expired test solution) implying that control solutions were not contaminated with Durad 310M

  

Table 3

 

Durad 310M- Determination of Acute Toxicity (EC50) to Daphnia (48 h, Semi-Static)

Cumulative Number of Immobile Daphnia Recorded During the Limit Test

  

Time (h)

Tank Replicate

Nominal Loading of Durad 310M (mg/l)

 

 

 

 

0

1000

24

I

0

0

II

0

0

III

0

0

IV

0

 

Per cent ImmobileDaphnia

0

0

 

 

 

 

48

I

0

1

II

0

0

III

0

0

IV

0

0

Per cent ImmobileDaphnia

0

5

Five Daphnia in each of four replicates were tested at each concentration

Validity criteria fulfilled:
yes
Conclusions:
This test was conducted on a sample of the material where the content of Triphenyl phosphate is present at <5%

The EL50 value of Durad 310M to Daphnia magna was determined to be >1000 mg/l based on initial loading rates and therefore not acutely toxic under the conditions of the test.
The No Observed Effect Loading (NOEL) for immobilisation was determined to be 1000 mg Durad 310M / L. The NOEL for observed effects is <1000 mg/l (<0.773 mg/l mean measured concentration).
Executive summary:

The acute toxicity of Durad 310M to Daphnia magna was assessed by conducting a limit test at a nominal loading rate of 1000 mg Durad 310M / l of medium. The study was conducted over a 48 h period under semi-static conditions, with test solutions being renewed at 24 h. The study was designed to comply with OECD (1984) Guideline 202 Part I.

 

Test solutions were prepared and tested as water accommodated fractions (WAF) from an initial loading rate of 1000 mg/l. Four test vessels at 1000 mg/l and four control vessels (0 mg/l) were included in the test, each vessel containing five Daphnia.The method of preparation was selected to ensure the test material was tested at the maximum achievable solubility under the conditions of the test.

 

Over the duration of the study one immobile Daphnia was observed at 1000 mg/l. This immobilisation is not regarded as significant. The results indicate that Durad 310M is not acutely toxic toDaphnia magna,under the conditions of the test. It is concluded that the EL50 of Durad 310M to Daphnia magna is >1000 mg/l under the test conditions. The No Observed Effect Loading (NOEL) for immobilisation is concluded to be equal to 1000 mg Durad 310M / l.

 

Water quality parameters (pH, temperature, conductivity and dissolved oxygen) were measured at the beginning and at 24 h intervals and remained within acceptable limits throughout the duration of the test.

On the basis of the study data, the substance is not Harmful to Daphnia.

Description of key information

Short-term toxicity to aquatic invertebrates

Key value for chemical safety assessment

EC50/LC50 for freshwater invertebrates:
1.5 mg/L

Additional information

Three studies are presented for acute toxicity to Daphnia Magna, with the following results:

 

Where TPP content > 5%

 

LC50 (48 h): 2.44 mg/L test mat. (nominal) based on: mortality  (K2)

LC50 (48 h): 1.5 mg/L test mat. (nominal) based on: mortality(K2)

 

Where TPP content < 5%

 

LC50 (48 h): > 1000 mg/L dissolved (nominal) based on: immobilisation (K1)

 

The data set available for the phosphates as a group indicates that these do pose hazardous effects to Daphnia.  However, such effects are considered to be attributable to the content of the impurity, triphenyl phosphate, CAS 115-86-6, (EC No. 204-112-2).  The content of this impurity is considered to adversely affect the toxicity of the substance within aquatic organisms. The registration dossier that two separate classifications applicable to the substance for this endpoint as follows:

 

• Phenol, isopropylated, phosphate (3:1) [Triphenyl phosphate >5%] - H410: Very toxic to aquatic life with long lasting effects.

 

• Phenol, isopropylated, phosphate (3:1) [Triphenyl phosphate < 5%] - H413: May cause long lasting harmful effects to aquatic life.

 

For the purposes of hazard classification, the following studies are taken as ket studies:

 

Where TPP content > 5%

LC50 (48 h): 1.5 mg/L test mat. (nominal) based on: mortality(K2)

 

Where TPP content < 5%

LC50 (48 h): > 1000 mg/L dissolved (nominal) based on: immobilisation (K1)