[[(phosphonomethyl)imino]bis[hexamethylenenitrilobis(methylene)]]tetrakisphosphonic acid

Sediment toxicity

Administrative data

Endpoint:

sediment toxicity: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

12/09/2002-26/09/2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to a test protocol that is comparable to the appropriate OECD test guideline/EU test method/national standard method, and in compliance with GLP. However, no analytical monitoring has been carried out.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Principles of method if other than guideline:

PARCOM Guideline: MAFF/ERT Harmonised Protocol. A Sediment Bioassay using an amphipod Corophium. Spp

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

[[(phosphonomethyl)imino]bis[hexamethylenenitrilobis(methylene)]]tetrakisphosphonic acid

EC Number:

252-156-6

EC Name:

[[(phosphonomethyl)imino]bis[hexamethylenenitrilobis(methylene)]]tetrakisphosphonic acid

Cas Number:

34690-00-1

Molecular formula:

C17H44N3O15P5

IUPAC Name:

[[(phosphonomethyl)imino]bis[hexamethylenenitrilobis(methylene)]]tetrakisphosphonic acid

Constituent 2

Reference substance name:

BHMT-H

IUPAC Name:

BHMT-H

Constituent 3

Reference substance name:

bis(hexamethylenetriaminepenta(methylene phosphonic acid)

IUPAC Name:

bis(hexamethylenetriaminepenta(methylene phosphonic acid)

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

SEDIMENT
- Concentrations: 50; 250; 875; 3000 and 10000 mg/kg (wet weight) of sediment
- Sampling interval: water quality (pH, DO and temperature) in the vessels was monitored three times during the 10 days of the test. Salinity was measured prior to the start of the test and on day 10 when the test was terminated.
- Sample storage before analysis: No analysis

PORE WATER: Not reported
- Concentrations:
- Sampling interval:
- Sample storage before analysis:

OVERLYING WATER: Not reported
- Concentrations:
- Sampling interval:
- Sample storage before analysis:

Test substrate

Vehicle:

no

Details on sediment and application:

PREPARATION OF SPIKED SEDIMENT
The top 5 to 10 cm of sediment was collected from an inter-tidal area free of contamination and transported to the laboratory in polythene bags. At the laboratory the sediment was sieved through a 500 µm sieve in a closed system using natural seawater and stored in sealed polythene bags in a dark room at 4 °C until required for use.
- Pooling or mixing of different substrates: The sediment was thoroughly homogenised immediately prior to use.
5 nominal concentrations (per unit wet weight) of spiked sediment were obtained as follows;
Approximately 450-500 g of wet sediment was placed in a 2 litre polypropylene container. One container per concentration and one for the control.The test substance was mixed with a small amount natural seawater before adding to the wet sediment.
A spatula was used initially to disperse the test substance throughout the sediment in the container and sufficient clean seawater was then added to create free flowing slurry.
- Method of mixing: The containers were then placed horizontally on a shaking table (approximately 100 revolutions per min.) for a minimum of 3 hours.
The following nominal wet weight concentrations were used in this test; 50; 250; 875; 3000 and 10000 mg/kg (wet weight) of sediment.
The containers were removed from the shaking table and shaken vigorously by hand for one minute immediately before the contents were dispensed to the appropriate replicate 1 litre tall-form glass test vessels.
The contents were dispensed in small aliquots to each replicate in turn to ensure an even chemical and particle size distribution.
- Equilibration time: The sediment was left to settle in the test vessels overnight or for a minimum of 3 hours. Within 24 hours of the sediment being introduced to the test vessel, natural seawater was carefully added to each vessel to bring the total volume up to the 900 ml mark. When the sediment had settled the vessels were aerated for 12 hours prior to the initiation of the test.


PREPARATION OF SPIKED WATER
- Details of spiking: Not stated
- Controls: Not stated

Test organisms

Test organisms (species):

Corophium volutator

Details on test organisms:

TEST ORGANISM
- Common name: Corophium volutator
- Strain/clone: Not specified
- Source: BioTrix in Trondheim, Norway, collected from Buvik in South Trondelag an area of inter-tidal mud/sand free of significant contamination.
- Details on collection: The test animals were sieved from the sediment 5 days before the initiation of the test and held in an aquarium at test conditions for acclimatisation.
- Age of animals at beginning of exposure: Adult 5mm or greater
- Feeding during test: Not specified
- Food type: Not specified
- Amount: Not specified
- Frequency: Not specified

ACCLIMATION
- Acclimation period: 5 days
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: Not specified
- Feeding frequency: Not specified
- Health during acclimation (any mortality observed): At AnalyCen Ecotox's laboratory a control chamber were set up to measure the mortality 5
days before testing. Mortality was 2 %.

Study design

Study type:

laboratory study

Test type:

static

Water media type:

saltwater

Type of sediment:

natural sediment

Limit test:

no

Exposure duration

Duration:

10 d

Exposure phase:

total exposure duration

Test conditions

Hardness:

Not reported

Test temperature:

Range 13.6-15.1 °C.

pH:

Range 8.0-8.1

Dissolved oxygen:

Range 98-100 %

Salinity:

Range 3.4-3.6

Ammonia:

Not reported

Nominal and measured concentrations:

Nominal: 50; 250; 875; 3000 and 10000 mg/kg (wet weight) of sediment.
No measured data

Details on test conditions:

TEST SYSTEM
- Test container (material, size): 1 litre tall-form glass test vessels
- Sediment volume: Not specified
- Weight of wet sediment with and without pore water: Not specified
- Overlying water volume: Not specified
- Depth of sediment and overlying water: 900 ml
- Aeration: yes
- Aeration frequency and intensity: Not specified
- Replacement of evaporated test water, if any: A constant water level (900 ml) was maintained in the test vessels by topping up with distilled water

EXPOSURE REGIME
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 3
- No. of replicates per control / vehicle control: 5
- Feeding regime: Not specified
- Type and preparation of food: Not specified
- Amount of food: Not specified

RENEWAL OF OVERLYING WATER
- Details on volume additions: Not specified
- Flow-rate: Not specified

OVERLYING WATER CHARACTERISTCS
- Type of water (e.g. deionized, ground water, sea water, Elendt medium acc. to OECD 219): Sea water. No other details specified

SOURCE OF NATURAL SEDIMENT
- Location and description of sampling site: Inter-tidal area
- Contamination history of site: Free of contamination

HANDLING OF NATURAL SEDIMENT
- Time of collection: Not reported
- Core depth: 5-10 cm
- Water depth: Not reported
- Storage conditions: The sediment was transported to the laboratory in polythene bags. At the laboratory the sediment was sieved through a 500µm sieve in a closed system using natural seawater and stored in sealed polythene bags in a dark room at 4 °C.
- Storage duration (prior to test): Not reported


OTHER TEST CONDITIONS
- Light quality: Dim lighting
- Photoperiod: Constant
- Light intensity: Dim

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
The salinity, temperature, pH and dissolved oxygen (DO) of the seawater in each vessel were measured immediately prior to initiating the test (day 0).
The behaviour of the Corophium was monitored throughout the test and the water quality (pH, DO and temperature) in the vessels was monitored on days 0, 1, 7 and 10 of the test.
Salinity was monitored immediately prior to the initiation of the test and the 10th day when the test was terminated.
On the 10th day the test was terminated by sieving the content of each beaker through a 500 µm sieve and counting the number of live and dead Corophium in each vessel. Missing animals were presumed to be dead and counted as such.

TEST CONCENTRATIONS
- Spacing factor for test concentrations:

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

10 d

Dose descriptor:

LC50

Effect conc.:

> 10 000 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 2

Duration:

10 d

Dose descriptor:

NOEC

Effect conc.:

877 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 3

Duration:

10 d

Dose descriptor:

NOEC

Effect conc.:

395 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Effect concentrations 4

Duration:

10 d

Dose descriptor:

LC50

Effect conc.:

> 4 500 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Reported statistics and error estimates:

LC50 calculated from a Logistic Regression Model.

Any other information on results incl. tables

Table 1: Test Concentrations and observed mortalities after 10 days for test substance DEQUEST 2090. Wet weight to dry weight ratio = 1.26

Test Concentration mg/kg dry weight	Log Values Test Concentration	No. Organisms per Vessel	Mortalities parallel 1	Mortalities parallel 2	Mortalities parallel 3	Mortalities parallel 4 (control only)	Mortalities parallel 5 (control only)
Control	-	10	0	0	0	1	2
50	1.7	10	0	0	0	-	-
252	2.4	10	0	1	0	-	-
877	2.9	10	0	0	0	-	-
3007	3.5	10	0	2	0	-	-
10013	4.0	10	3	4	4	-	-

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

A 28 day LC50 value of >10000 mg/kg dw sediment and a NOEC value of 877 mg/kg dw sediment have been reported for the effects of the test substance on the mortality of the marinewater amphipod Corophium volutator. The results are based on nominal concentrations.