Phosphinic acid, P-[3-(acetyloxy)-3-cyanopropyl]-P-methyl-, butyl ester

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:

1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1930 (Mysid Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations, all replicates at test initiation and termination
- Sampling method: Samples
taken on Day 0 were acidified with 5 drops of phosphoric acid to an approximate pH of 3.0
- Sample storage conditions before analysis: A sample of the 10 mg/L target nominal (7.75 mg/L actual nominal) test solution was collected and acidified with 5 drops of phosphoric acid to an approximate pH of 3.0. The sample was stored at room temp in the analytical lab.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The targeted nominal test concentrations were 0.625,1.25, 2.5, 5, and 10 mg/L. However, The measured stock solution concentration was 16.74 mg/L instead of the 21.6 mg/L target nominal stock
concentration. Therefore the actual nominal values, based on addition of the measured stock solution were 0.484, 0.969,1.94, 3.88, and 7.75 mg/L.
Test solutions were delivered to duplicate test chambers per
concentration via a proportional diluter.

Test organisms (species):

Americamysis bahia (previous name: Mysidopsis bahia)

Details on test organisms:

TEST ORGANISM
- Justification for species other than prescribed by test guideline:
- Age at study initiation (mean and range, SD): less than 24 h
- Weight: mean of control mysids 0.107 mg (end of the study)
- Stage and instar at study initiation: juvenile
- Method of breeding:
Temperature: 24.8 to 25.3 °C
Light: 14 hours light and 10 hours darkness
Food: Concentrated Artemia nauplii
Vessels: Brood Stock: 36.9 liter glass fish tank, containing approximately 28.9 liters of dilution water
Water: Synthetic sea water in a recirculating system
- Source: Gravid female mysids (Lot M-20) were purchased from Chesapeake Cultures, Hayes, VA, juvenile in house
- Feeding during test: yes
- Food type: concentrated Artemia nauplii
- Frequency: twice daily

ACCLIMATION
- Acclimation period: > 24 h brood stock
- Type and amount of food: concentrated Artemia nauplii
- Health during acclimation (any mortality observed): no mortality observed

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS:
Juveniles: Collection chamber, made up of a Pyrex® crystallizing
dish 125 mm diameter by 65 mm high, with an 18 cm high 300 micron square nitex screen, placed inside a 19.5 liter glass fish tank, containing approximately 13 liters of dilution water

Test type:

flow-through

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

Temperature: 24.8 to 25.3 °C

pH:

see table 1

Dissolved oxygen:

> 95 % saturation

Salinity:

ca. 20 %

Nominal and measured concentrations:

Nominal: 0.484, 0.969, 1.94, 3.88, and 7.75 mg/L
Measured: 0.46, 0.90, 1.71, 3.34, 6.40

Details on test conditions:

TEST SYSTEM
- Test vessel: 1 liter Pyrex® beakers with a 9.0 cm high 300 micron square nitex screen
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 10.2 cm in diameter, and
15.5 cm high, with a test solution depth of approximately 15.5 cm
- Volume of solution: 1.2 L
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate):
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- Biomass loading rate: 0.892 mg/L

TEST MEDIUM / WATER PARAMETERS
- Preparation of dilution water: synthetic sea water, prepared by
adding synthetic sea salts
- Parameters: bi- weekly monitoring, see table 1 for results
- Intervals of water quality measurement: broad range of contaminants and water quality parameters = twice a year; bi-weekly monitoring for nitrite, ammonia, salinity, pH,
and dissolved oxygen

OTHER TEST CONDITIONS
- Photoperiod: 14:10, light:dark with gradual intensity changes at dawn and dusk
- Light intensity: 116 foot candles

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Observations for death, and for abnormal appearance and behavior, were performed at 24, 48, 72, and 96 hours (± 1 hour).

RANGE-FINDING STUDY
- Test concentrations: 0.5,1.0,5.0, and 10 mg/L (nominal)
- Results used to determine the conditions for the definitive study: LC50 (72h) = 1.4 mg/L, no additional mortalities between 72 and 96 h

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

3.62 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CL: 1 .71 to 6.40 mg/l

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

2.57 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CL: 1.71 to 3.34 mg/l

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

1.88 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CL: 1.57 to 2.25 mg/l

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.79 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% CL: 1.52 to 2. 13 mg/l

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

0.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

The 96 hour LC50 of the test substance to the mysid (Mysidopsis bahia) under the conditions of this study is 1.79 mg/L (slope = 6.9). The no observed effect concentration (NOEC) is 0.46 mg/L. The lowest observed effect concentration (LOEC) is 0.90 mg/L.
There was one random mortality, and no abnormal behavior or appearance in the control mysids during the study.

Validity criteria fulfilled:

yes

Conclusions:

The test material is toxic to the mysid Mysidopsis bahia under the conditions of this study.

Executive summary:

An experimental study according to the guideline EPA OTS 797.1930 and GLP regulations was conducted to assess the acute toxixity of the test item to marine aquatic invertebrates. Mysidopsis bahia was exposed to the nominal concentrations 0.484, 0.969, 1.94, 3.88, and 7.75 mg/L in a flow through system for 96 h. 10 juvenil mysids (under 24 h old) were randomly picked from the  acclimated stock culture for each test vessel. Duplicates were used for every concentration. After 96 h an EC50 of 1.79 mg/L was determined and a NOEC of 0.46 mg/L.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: EPA; - Methods for Acute Toxicity Tests with Fish, Macroinvertebrates, and Amphibians (EPA-660/3-75-009) and - Pesticides Assessment Guidelines, Subdivision E, §72-2: Acute toxicity test for freshwater aquatic invertebrates.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: start and at the end of testing from all tested nominal concentrations

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
Method: test substance was weighed out to a precision of 0.1 mg and mixed in reagent-grade water as a stock solution. Oefined amounts of stock solution - or dilutions of it - were pipetted
proportionally into the filled jars and stirred vigorously with a glass rod. 2000 ml test water were prepared in volumetric flasks for all tested concentrations

- Other relevant information: Calculations of the aqueous concentrations of test substance were based on 100% purity of the test material

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: STRAUS, Clone 5 (Bradley, 1988)
- Age at study initiation: 1- 24 h
- Stage and instar at study initiation: juvenil
- Method of breeding:
Cultured in beakers (19x13 cm), containing initially 20 adult animals in 2 litre test water.
Approximately twice a month the daphnids were transferred into fresh water resulting in the initial biological loading.
The breeding temperature was 20 ± 2°Celsius.
Food: cultures of monocellular green algae (Scenedesmus subspicatus CHODAT, strain 86.81 SAG
The algae were separated from the nutrient medium, suspended in filtered water, and served twice a week.
Water used for breeding was the same as used in the test.
- Source: Hoechst Schering AgrEvo GmbH, Umweltforschung, Oekobiologie, D-65926 Frankfurt am Main, Germany
- Feeding during test: no

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES, INCLUDING CULTURING CONDITIONS:
The gravid mother animals were separated from the culture by means of a glass tube and then transferred into a 2 L crystallising dish, filled with test water.
After a maximum of 24 hours the new-born daphnids were sampled at random with a glass tube.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

32.3 mg/L as CaCo3

Test temperature:

mean = 19.9 °C(see table 1)

pH:

mean = 7.8 (see table 1)

Dissolved oxygen:

mean = 8.8 mg/L (see table 1)

Conductivity:

mean = 148 µS/cm (see table 1)

Nominal and measured concentrations:

Nominal (test material): 3.2, 5.6, 10, 18, 32, 56, 100 and control;

Nominal (active ingredients): 2.829, 4.95, 8.84, 15.912, 28.288, 49.504, 88.4 and control;

Mean measured (active ingredients): 2.38, 4.54, 8.60, 13.60, 24.59, 50.46, 88.51 and control

Details on test conditions:

TEST SYSTEM
- Test vessel: 300 ml glass jars
- Type (delete if not applicable): closed with glass lids
- Size: 95 mm and a height of 50 mm. water depth = 30 - 31 mm
- Volume of solution: 200 ml test water
- Aeration: no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): For the concentrations of 3.2, 5.6, 10, 18, 32, 56, 20 daphnids in 2 beakers were tested.
For the concentration of 100 mg/L 60 daphnids in 6 beakers were tested.
- No. of vessels per control (replicates): 2
- other: additional flask for chemical analysis were prepared with organisms

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionized water, additionally filtered by a deionization unit and a charcoal filter within the laboratory before use.
- Medium: soft artificial mineral medium M4 (Elendt 1990), slightly modified. To prevent precipitation during preparation of medium, the chemicals were given into 500-800 mL deionized water and then filled up to 1000 ml
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16:8 h light:dark
- Light intensity: wide spectrum fluorescent lights

EFFECT PARAMETERS MEASURED:
immobilization and intoxication symptoms were observed after 24 and 48 h

Reference substance (positive control):

no

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

11.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 9.5 - 14.1 mg/L

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

9.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 7.8 - 10.7 mg/L

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

5.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Evaluations
Test criterion was the immobilisation of animals. The animals were counted as immobile, when they were not able to swim within 15 seconds after gentle agitation of the test container.

- Other biological observations: lntoxication symptoms were not observed during the times of observations
- Mortality of control: none
- Other adverse effects control: none
- Immobilisation of control: none

Reported statistics and error estimates:

The concentration of test substance estimated to immobilize 50% of the test population (EC50) with the 95% confidence limits was determined for each evaluation time using a computerized calculation program (Stephan, C.E., 1982). This program employs the binomial probability, moving average and probit method for the calculation. The results of the method giving the
narrowest confidence limits for the EC50 was selected for this report.
At the end of testing the concentration-effect relationship was plotted for the 24 h determination using a computer program (Microsoft® Excel, Vers. 5.0).
Mean values with standard deviation were determined for all measured physical and chemical parameters.

Validity criteria fulfilled:

yes

Conclusions:

The substance is toxic to dahnids under the conditions of this study.

Executive summary:

An experimental study according to the guideline EPA-660/3-75-009 and GLP regulations was conducted to assess the acute toxicity of the test item to aquatic invertebrates in freshwater. Daphnia magna was exposed to 3.2, 5.6, 10, 18, 32, 56, 100 mg/L (nominal concentration, based on test material) for 48 h in a static system. The measured concentrations were 2.38, 4.54, 8.60, 13.60, 24.59, 50.46 and 88.51 mg/L (based on the active ingredients). Per test vessel 10 daphnids in the age of 0-24 h (initial age) were exposed in two replicates. Only for the highest concentration 6 vessels with 10 daphnids were used. For the measurements of physical and chemical parameters one replicate was prepared of the untreated control, the high, medium and low concentrations in order to avoid stress to the daphnids by performing the measurements. After 48 h an EC50 of 9.1 mg/L and a NOEC of 5.6 mg/L was determined. No unforeseen circumstances were observed which may have affected the quality or integrity of this study and the validity criteria were fullfilled.

Description of key information

The test item is acutely toxic for marine and freshwater aquatic invertebraes. The lower EC50 for marine organisms indicates a higher toxixity of the test item to marine invertebrates.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

9.1 mg/L

Marine water invertebrates

Marine water invertebrates

Dose descriptor:

EC50

Effect concentration:

1.79 mg/L

Additional information

An experimental study according to the guideline EPA-660/3-75-009 and GLP regulations was conducted to assess the acute toxicity of the test item to aquatic invertebrates in freshwater. Daphnia magna was exposed to 3.2, 5.6, 10, 18, 32, 56, 100 mg/L (nominal concentration, based on test material) for 48 h in a static system. Per test vessel 10 daphnids in the age of 0-24 h (initial age) were exposed in two replicates. Only for the highest concentration 6 vessels with 10 daphnids were used. For the measurements of physical and chemical parameters one replicate was prepared of the untreated control, the high, medium and low concentrations in order to avoid stress to the daphnids by performing the measurements. After 48 h an EC50 of 9.1 mg/L and a NOEC of 5.6 mg/L was determined. No unforeseen circumstances were observed which may have affected the quality or integrity of this study and the validity criteria were fullfilled (Hoest AG, 1996).

To assess the acute toxixity of the test item to marine aquatic invertebrates, an experimental study according to the guideline EPA OTS 797.1930 and GLP regulations was conducted  Mysidopsis bahia was exposed to the nominal concentrations 0.484, 0.969, 1.94, 3.88, and 7.75 mg/L in a flow through system for 96 h. 10 juvenil mysids (under 24 h old) were randomly picked from the  acclimated stock culture for each test vessel. Duplicates were used for every concentration. After 96 h an EC50 of 1.79 mg/L was determined and a NOEC of 0.46 mg/L.

These resluts indicate an acute toxicity of the test item to aquatic invertebrates in freshwater and marine systems.