[No public or meaningful name is available]

Sediment toxicity

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

sediment toxicity: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2019-12-02 to 2019-12-16, with the definitive exposure phase from 2019-12-03 to 2019-12-13.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: DIN EN ISO 16712

Version / remarks:

2006

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Determination of the test item
The concentration of the test item was analytically verified via LC-MS/MS. Recoveries below 80% were expected at day 10 analysis in the limit concentration, due to non-extractable test item (i.e. bound residues) left in the sediment. Therefore, analysis was carried out on the day of application of the test item (day -1), on day of test organism insertion (start of the exposure, day 0) and at the end of the exposure (day 10). On the day of application (day -1), the stock solution, the spiked quartz sand and the sediment were analytically verified after preparation. At the start (day 0) and at the end of the exposure (day 10), the test item was analytically verified in the water layer as well as in the sediment layer. Two additional replicates of the limit concentration, the solvent control and the control for the test item analysis were prepared without animals and incubated under test conditions. Water samples and sediment samples were separated by decantation. After decantation of the remaining water phase, the test item was extracted from the sediment and analyzed. The samples were analyzed with an LC-MS/MS method, implemented under non-GLP and documented finally in the GLP raw data

Vehicle:

no

Details on sediment and application:

Dry weight of the test sediment
Before experimental starting, the dry weight of the test sediment was determined to be 72%.


Stock solution
For the limit test item concentration, a stock solution was prepared with the solvent acetone (see below) one day before the start of the exposure (day -1).
For the preparation of the stock solution, an appropriate amount of the test item was weighed out, dissolved in an appropriate amount of acetone and treated with ultrasound for 15 minutes at room temperature. The nominal concentration of the stock solution was calculated in relation to the dry weight and the amount of the test sediment. For preparation of 1500 g sediment with a dry weight content of 72%, a stock solution of 36 g/L in acetone (1800 mg test item / 50 mL) was prepared.

Limit concentration
A limit concentration of 796 mg test item/kg sediment (dry weight) was prepared and tested.
Application 30 mL of the acetone stock solution mentioned above were mixed with 60 g of quartz sand and the solvent was evaporated completely by air under the laboratory fume hood. 1500 g of the natural wet test sediment (dry weight: 1080 g) were mixed with 50 g of the spiked quartz sand for at least 3 minutes.

Control
Natural sediment from the collecting site of the amphipods with dilution water (both without test item) was tested under the same test conditions as the test groups.

Solvent Control
Natural sediment from the collecting site of the amphipods mixed with an appropriate amount of quartz sand, which was treated with acetone (same procedure as described above for the limit concentration), and overlaid with dilution water (all without test item) was used as a solvent control and was tested under the same conditions as the test and control replicates.

Solvent
Acetone
[supplier: VWR, batch: 190325A001, purity: ≥ 99.90% (Pestinorm® for GC, expiry date: 03/2022]

Test organisms (species):

Corophium volutator

Details on test organisms:

Test system
Corophium volutator PALLAS 1766
(Arthropoda, Diantennata, Crustacea, Malacostraca, Amphipoda, Gammaridea, Corophiidae)

Reason for the selection of the test system
Corophium volutator, a marine sediment dwelling amphipod species was used according to the demands of the test guideline.

Origin
The animals were collected from a clean intertidal shore (mud or muddy sand) known to be free from contamination (Norderney, Surfbucht, North Sea, Germany) and kept in the laboratory under suitable holding conditions as specified below. The collection area is accepted by German national authorities and known to be free from pollution.
All amphipods used in the test originated from the same collecting site and collecting date.

Holding / Acclimation under test conditions
Since the day of collection (2019-11-18), the amphipods were held at the facility until the start of the study (15 days after collection) in glass aquaria with filter system, filled with natural sediment from the collecting site (actual depth: 3 - 4 cm) and overlying water (actual depth: approximately 15 - 16 cm) with a salinity of 28 +/- 2 S (~28 +/- 2 g/kg), a pH value of 8 +/- 0.5 at 15 +/- 2 °C and diffuse light (500 - 1000 lx, photoperiod 16 hours daily).
The dissolved oxygen concentration of the overlying water was ≥ 85% of the air saturation value.
The animals were cultivated with a density of one animal per cm². Any individuals that failed to bury in holding sediment or that emerged from the sediment at any time during the holding / acclimation period and appeared dead or inactive when gently prodded or that appeared unhealthy during holding were discarded.

Holding / overlying water
Reconstituted salt water was used. It was prepared by adding an appropriate amount of commercially available sea salt (SIGMA) to demineralized water to achieve a salinity of 28 +/- 2 S.
The reconstituted salt water was allowed to stand for 1 - 2 weeks before use. The overlying water was aerated in the holding / test containers. The dissolved oxygen content of the seawater overlying the sediment was close to the air-saturation value during the test organism holding or acclimation period, at test initiation and throughout the 10-days test period.

Holding and test sediment
atural sediment from the collecting site of the amphipods was used after removal of coarse particles and other sediment dwelling organisms. For this purpose, the sediment was sieved after receipt with a nylon-mesh sieve (sieve size 0.5 mm).
Dry weight, concentration of ammonia and sulphide, pH-value and salinity of the pore water of the test sediment were determined.

Feeding
Food was provided twice per week ad libitum. The test amphipods were not fed within 3 days before the start of the exposure phase of the test.

Food
A suspension of the marine Diatomae Phaeodactylum tricornutum was provided as food. The algae are cultured at the test facility.

Study type:

laboratory study

Test type:

static

Water media type:

saltwater

Type of sediment:

natural sediment

Limit test:

yes

Duration:

10 d

Exposure phase:

total exposure duration

Hardness:

sse any other information on materials and methods incl. tables

Test temperature:

sse any other information on materials and methods incl. tables

pH:

sse any other information on materials and methods incl. tables

Dissolved oxygen:

sse any other information on materials and methods incl. tables

Salinity:

sse any other information on materials and methods incl. tables

Ammonia:

sse any other information on materials and methods incl. tables

Conductivity:

sse any other information on materials and methods incl. tables

Nominal and measured concentrations:

A limit concentration of 796 mg test item/kg sediment (dry weight) was prepared and tested.

Details on test conditions:

Test method
Static

Test duration
10 days

Test vessels
Glass beakers (9 (ID) x 18 (H) cm), 1000 mL capacity, loosely covered with polyethylene tops

Number of replicates
3 replicates of the limit concentration, the solvent control and the control (with amphipods) and 2 separate replicates for the analytical monitoring (without amphipods)

Number of amphipods per replicate
20 amphipods per replicate


Test volume
ca. 900 mL per replicate (total):
ca. 175 - 200 mL sediment, corresponding to a layer of 3 cm and
ca. 700 mL of the test solution / overlying water

Dilution water
Same as holding / overlying water

Sediment
Sieved sediment taken from the collecting site of the amphipods (same as holding sediment)

Preparation of test vessels
After completion of the mixing period of the sediment with the spiked quartz sand, equal aliquots of the mixtures were filled into the test vessels (278 g sediment were weighed into each test vessel). The sediment layer had a depth of approximately 3 cm. The overlying water was filled up to a total volume of 850 mL in the test vessel with the respective test solutions and aerated approximately 24 hours before start of the exposure.

Aeration
Gentle aeration was provided throughout the test.

Age of amphipods at the start of the exposure
Adult Corophium volutator, not greater than approximately 12 mm in length (without antennae)

Introduction of amphipods
Amphipods were sieved (sieve size 0.5 mm) from the holding sediment. Active, apparently healthy amphipods were selected randomly and sequentially distributed among sorting beakers containing approximately 50 mL dilution water with a featherweight forceps until each beaker contains 20 individuals. Amphipods were introduced to the test vessels without disruption of the sediment by gently pouring the water and amphipods from the sorting beakers into the test vessels.
No amphipods were replaced, because all amphipods had burrowed into the sediment within 1 hour in the solvent control and in the control. Several amphipods in the limit concentration showed an avoidance behavior to the test substrate during the first days and were not replaced.
 
Test temperature (target)
15 ± 2 °C (water temperature)

Dissolved oxygen concentration
Not less than 85% of air saturation value in the overlying water.


pH value
8 ± 0.5

Illumination (target)
Diffuse light, light intensity of 500 – 1000 lx

Photoperiod
Continuously illumination was provided to increase the tendency of the amphipods to remain buried in the sediment and to prevent nocturnal migration of the amphipods into the water phase.

Feeding
The amphipods were not fed during the test.

Duration:

10 d

Dose descriptor:

NOEC

Effect conc.:

>= 796 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

10 d

Dose descriptor:

LC10

Effect conc.:

> 796 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

10 d

Dose descriptor:

LC50

Effect conc.:

> 796 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

10 d

Dose descriptor:

LC100

Effect conc.:

> 796 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Reported statistics and error estimates:

Statistical evaluation
The NOEC for the mortality was determined by statistical evaluation as specified below. Significant deviations were determined in comparison to the control using statistical standard procedures as normality test, equal variance test and analysis of variance (i.e. parametric ANOVA).
Prior to running the analysis of variance, a normality test and an equal variance test was performed. P-values of the normality and equal variance tests were 0.05. The -value (acceptable probability of incorrectly concluding that there is a difference) was =0.05.
The determination of significant mortality was conducted with ANOVA One Way Analysis of Variance (P < 0.05).
The statistical evaluation was carried out with the software SigmaPlot for Windows, Version 14.0. Details of the statistical analyses.
All effect values given are based on the nominal concentration of the test item in the sediment layer.

Methods of evaluation
Due to the limit test design, the LC10 / 50 / 100-values (after 10 days) were estimated directly from the observation data, without mathematical calculation.

Software
All data are computer processed and rounded for presentation. Consequently, minor variations may occur from the original figures if manual calculations based on the original figures are made subsequently. Calculations were made using the following software:
- SigmaPlot for Windows, Version 14.0, Systat Software, Inc.
- Excel, MICROSOFT CORPORATION

Mortality in [%] after 10 days of Exposure in the Definitive Test

      (n = 60, divided into 3 replicates with 20 amphipods each)

Nominal

test item concentration       MORTALITY [%] after 10 days

      Replicates

[mg/kg sediment dry weight]       1       2       3       Mean

796       0       0       0       0

Solvent control       5       0       5       3

Control       0       5       0       2

Absolute Numbers of dead Amphipods after 10 days of Exposure in the Definitive Test

                    (n = 60, divided into 3 replicates with 20 amphipods each)

Nominal

test item concentration       Number of dead amphipods

[N]       Number of amphipods

which were not found

[N]       Total number

of amphipods

[N]

      After 10 days

      Replicates              

[mg/kg sediment dry weight]       1       2       3       Mean       1       2       3       Σ       per

replicate

796       0       0       0       0       all found       all found       all found       60       20

Solvent control       1       0       1       2       all found       all found       all found       60       20

Control       0       1       0       1       all found       all found       all found       60       20

 Additional Observations during the Definitive Test

The amphipods were buried within 1 hour after insertion in all replicates of the solvent control and the control. After 24 hours, 63% of the inserted amphipods were swimming in the water layer at the limit concentration of 796 mg/kg sediment dry weight. During the first days, the amphipods avoided burying into the sediment at the limit concentration. The daily observations are presented in Table 4. Therafter, the amphipods were mostly buried. The sediment of the limit concentration was black and had a strong sulfic odur of H2S at the end of the exposure period, whereas the sediment in the solvent control and the control was brownish and had no abnormal odor.

  Additional Observations during the 10-days of Exposure Period in the Definitive Test

                    (n = 60, divided into 3 replicates with 20 amphipods each)

Nominal test item concentration

[mg/kg  sediment dry weight]       Control       Solvent control       796

Day

      Date

(yyyy-mm-dd)       Parameter

      Replicates

                    1       2       3       1       2       3       1       2       3

 0       2019-12-03

(after 1 hour)       A       20       20       20       20       20       20       11       7       4

             D       0       0       0       0       0       0       9       13       16

 1       2019-12-04       A       20       20       20       20       20       20       7       10       5

             D       0       0       0       0       0       0       13       10       15

 2       2019-12-05       A       20       20       20       20       20       20       10       16       13

             C       0       0       0       0       0       0       2       3       6

             D       0       0       0       0       0       0       8       1       1

 3       2019-12-06       A       20       20       20       20       20       20       10       16       15

             C       0       0       0       0       0       0       0       3       1

             D       0       0       0       0       0       0       10       1       4

 4       2019-12-07       A       20       20       20       20       20       20       10       16       13

             C       0       0       0       0       0       0       9       3       4

             D       0       0       0       0       0       0       1       1       3

 5       2019-12-08       A       20       20       20       20       20       20       11       13       16

             C       0       0       0       0       0       0       7       7       1

             D       0       0       0       0       0       0       2       0       4

 6       2019-12-09       A       20       19       20       20       20       20       15       16       11

             B       0       1       0       0       0       0       0       0       0

             C       0       0       0       0       0       0       4       3       7

             D       0       0       0       0       0       0       1       1       2

 7       2019-12-10       A       20       19       20       20       20       19       18       17       13

             C       0       0       0       0       0       1       1       2       2

             D       0       0       0       0       0       0       1       1       5

 8       2019-12-11       A       20       19       20       20       20       20       14       19       16

             C       0       0       0       0       0       0       5       1       2

             D       0       0       0       0       0       0       1       0       2

 9       2019-12-12       A       20       19       20       20       20       19       19       18       18

             B       0       0       0       0       0       1       0       0       0

             C       0       0       0       0       0       0       1       2       2

10       2019-12-13       A       20       18       20       19       19       19       20       20       18

             C       0       1       0       0       0       0       0       0       0

             D       0       0       0       0       1       0       0       0       2

             E       0       0       0       1       0       0       0       0       0

A       = number of amphipods buried

B       = number of dead amphipods on the sediment layer

C       = number of living amphipods on the sediment layer

D       = number of amphipods swimming in the water layer

E       = number of dead amphipods in the sediment layer

Water Quality Parameters

The measured water quality parameters (i.e. pH-value, dissolved oxygen concentration and salinity, concentrations of ammonia and sulphide in the pore water) were within the acceptable limits.

During the test period, the water temperature was recorded. Additionally, the room temperature in the climatic chamber was recorded and was within the range of 13.5 – 15 °C.

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentration of the test item Reaction products of DL-methionine and C18 unsaturated fatty acid chloride and isopropanol, the LC50 for mortality of Corophium volutator PALLAS was > 796 mg/kg sediment dry weight. The NOEC after 10 days was ≥ 796 mg/kg sediment dry weight.

Executive summary:

In the acute toxicity test with the amphipod Corophium volutator PALLAS in a water-sediment system, the effects of the test item Reaction products of DL-methionine and C18 unsaturated fatty acid chloride and isopropanol (batch no. ESD0033040) were determined at the test facility according to DIN EN ISO 16712 (2006) from 2019-12-02 to 2019-12-16, with the definitive exposure phase from 2019-12-03 to 2019-12-13.

A limit test under static conditions was conducted by spiking the sediment layer. A nominal test item concentration of 796 mg/kg sediment dry weight was tested. Since the limit concentration had to be prepared by using the solvent acetone, a solvent control and an untreated control were included in this study. Sixty amphipods, divided into 3 replicates with 20 amphipods per replicate, were exposed each to the limit concentration, the solvent control and the control.

The test item is a brownish liquid which is partially soluble in water. For preparation of the limit concentration, a stock solution of the test item of 36 g/L was prepared in the solvent acetone (Pestinorm for GC, ≥ 99.90%, VWR). The acetone stock solution was mixed with 60 g of quartz sand and the solvent was evaporated completely by air under the laboratory fume hood. 1500 g of the natural wet test sediment (dry weight content: 72%) was mixed with 50 g of the spiked quartz sand for at least 3 minutes. The sediment for the solvent control was prepared equivalently without test item.

After preparation of the sediment, 278 g of the natural wet test sediment (dry weight: 200 g) were filled into each test vessel, giving a sediment layer of approximately 3 cm depth in each test replicate. The sediment layer was overlaid with 650 mL of reconstituted salt water (28 ± 2S, SIGMA). The test vessels were aerated approximately 24 hours before insertion of the amphipods. The amphipods were introduced into the test vessels by gently pouring the water (50 mL) containing 20 amphipods from a sorting beaker into a test vessel.

The amphipods were buried within 1 hour after insertion in all replicates of the solvent control and the control. After 24 hours, 63% of the inserted amphipods were swimming in the water layer at the limit concentration of 796 mg/kg sediment dry weight. During the first days, the amphipods avoided burying into the sediment at the limit concentration. For details, see Table 4. Therafter, the amphipods were mostly burried. The sediment of the limit concentration was black and had a strong sulfic odur of H2S at the end of the exposure period, whereas the sediment in the solvent control and the control was brownish and had no abnormal odor.

The test item did not induce statistically significant mortality in the limit concentration, the solvent control and the control during the test period of 10 days (One Way Analysis of Variance, P < 0.05). The observed mortality was 2% in the control, 3% in the solvent control and 0% in the limit concentration of 796 mg/kg sediment dry weight.

Based on the nominal concentration of the test item Reaction products of DL-methionine and C18 unsaturated fatty acid chloride and isopropanol, the LC50 for mortality of Corophium volutator PALLAS was > 796 mg/kg sediment dry weight. The NOEC after 10 days was ≥ 796 mg/kg sediment dry weight.

Samples of the stock solution of the test item Reaction products of DL-methionine and C18 unsaturated fatty acid chloride and isopropanol, the spiked quartz sand and the sediment were analytically verified via LC MS/MS directly after preparation or application (day -1).

Samples of the water layer and the sediment layer (spiked with the test item) from an additionally prepared test replicate were analytically verified via LC-MS/MS on the day of test organism insertion (start of the exposure, day 0) and at the end of the exposure (day 10) of the limit concentration, control and the solvent control.

The measured concentration of the test item in the stock solution on day -1 was 93% of the nominal value. The measured concentration of the test item in the quartz sand on day -1 was 88% of the nominal value. The measured concentration of the test item in the water layer was < 1% of the nominal value at the start (day 0) and < LOQ at the end of the exposure (day 10). In the sediment layer, the measured concentration of the test item was 93% of the nominal concentration on day -1, 84% of the nominal concentration at the start of the exposure (day 0) and 27% at the end of the exposure (day 10).

Procedural recovery was checked via quality controls of the test item for the water phase and sediment at the lowest fortification level, prepared on each sampling date and treated in parallel to the test sample. Recovery rates were > 90% of the nominal concentration, fulfilling the criterion for the accuracy.

LC10 / 50 / 100 and NOEC

      (based on the nominal concentration of the test item)

Reaction products of DL-methionine and C18 unsaturated fatty acid chloride and isopropanol

Effect levels

      Toxicity endpoint

based on the nominal concentration of the test item

[mg/kg sediment dry weight]

LC10       > 796

LC50       > 796

LC100       > 796

NOEC1)       ≥ 796

1)       = One Way Analysis of Variance, P < 0.05

Reference 2

Endpoint:

sediment toxicity: short-term

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

08.03.07 -26.03.07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: PARCOM 1995 Part A

Principles of method if other than guideline:

PARCOM 1995 Part A: a sediment bioassay using an amphipod Corophium sp. (as described
in SOP no. 906)
Adult Corophium volutator (Pallas), a sediment-dwelling amphipod, of a size 5-10 nun are exposed to
spiked sediment for a period of 10 days. Water quality is recorded at specified intervals during the test.
At the end of the experiment the animals are sieved out of the sediment and the number of surviving
animals recorded. Survival is analysed statistically and compared with a control (non-spiked)
sediment. The objective is to determine the concentration of test item that in 10 days kills 50% of the
exposed test animals (10-day LCso).

GLP compliance:

yes

Specific details on test material used for the study:

Test item name:
Batch/Ref. ID No.:
Manufacturer/Supplier:
Date test item received:
Study Ref. No.:
Dodicor V 5654
07-YS-019
C1ariant GmbH
12.02.07
2221 \corophium

Analytical monitoring:

yes

Vehicle:

no

Details on sediment and application:

The sediment used for the test described in this report can be classified as a muddy, fine sand, with
particle characteristics approximating those recommended in the test guideline. Sediment samples are
obtained from sites supporting viable populations of Corophium volutator.
The wet to dry weight ratio of the sediment is determined by drying a sample in a tared cmcible at
60°C for 24 hours and reweighing after cooling to room temperature. Nominal test concentrations in
milligrams per kilogram dry sediment weight are determined from the wet to dry weight ratio.
The process of chemically spiking the sediment is intended to ensure that the test item is evenly
distributed throughout the test sediment, and that adequate contact between the test item and sediment
is attained.
Note: the method does not purport to reproduce the adsorption characteristics that may obtain under
equilibrium conditions or as a consequence ofchronic exposure in the field.
Before preparation of spiked sediment samples the test item is characterised as follows:
• Materials with a log Pow >4.0 or test items containing components with log Pow >4.0 are
initially dissolved in a low toxicity solvent (usually acetone) before addition to a small
quantity of dried sediment. Test items with an OSPAR behaviour category Ic, 2, 3 or 4 (see
below) are coated onto dried sediment before addition to the main mass of wet sediment.
• Materials or test items known to be soluble or dispersible in water are mixed with a small
quantity of seawater before direct addition to the wet base sediment.
• Powders are dissolved or dispersed in an appropriate medium before addition to either dried or
wet sediment.
In cases where the properties of chemicals or products are not clearly identified by prior information,
an examination of the behaviour of the test item in seawater is performed in accordance with the
OSPAR Guidelines for Toxicity Testing of Substances and Preparations Used and Discharged
Offshore.

Test organisms (species):

Corophium volutator

Details on test organisms:

Corophium volutator and their native sediment are collected locally. Animals are sieved from the
sediment on site and kept in 20-1 glass tanks of ambient seawater, with available food (detritus
collected with animals), for a period of acclimation to test conditions. Test sediment is sieved (500
).lm) in the laboratory using reference seawater to remove indigenous Corophium volutator or potential
predator organisms, and to adjust interstitial salinity to that of the reference seawater. The sieved
sediment is kept at 5°C in 30-1 covered plastic containers until use. Any supernatant water is decanted
and discarded. The quality (pH, dissolved O2, temperature and salinity) of the seawater in which the
Corophium volutator are transported is recorded on arrival at the laboratory. If necessary, the animals
are acclimated to the salinity of the test seawater by adjustment of the salinity of the holding water at a
rate not exceeding 4 mS.cm't per day. Animals are also acclimated to the test temperature (15 ± 2°C).

Study type:

laboratory study

Test type:

static

Water media type:

saltwater

Type of sediment:

natural sediment

Limit test:

no

Duration:

10 d

Exposure phase:

total exposure duration

Test temperature:

15 +- 2 °C

pH:

7.39 - 8.16

Dissolved oxygen:

85 - 99 % O2

Salinity:

natural seawater ca. 39 mS(cm / 25 Promille

Nominal and measured concentrations:

164 - 528 - 1634 - 5154 - 16654 mg/kg sediment and a control

Duration:

10 d

Dose descriptor:

LC50

Effect conc.:

7 461 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Validity criteria fulfilled:

yes

Conclusions:

An assessment ofthe solid-phase (sediment) toxicity (lO-day LCso) of Dodicor V 5654 was made
using the sediment-dwelling amphipod, Corophium volutator (Pallas). The following result, expressed
as mg Dodicor V 5654 per kg dry sediment weight, was obtained:
10-day LC50: 7461 mgkg-1 (95% c.i. 3288 - 14821)

Executive summary:

An assessment ofthe solid-phase (sediment) toxicity (lO-day LCso) of Dodicor V 5654 was made

using the sediment-dwelling amphipod, Corophium volutator (Pallas). The following result, expressed

as mg Dodicor V 5654 per kg dry sediment weight, was obtained:

10-day LC50: 7461 mgkg-1 (95% c.i. 3288 - 14821)

Comments: Mortality in the controls at the end of the test was 30%. Mortality in test vessels was

corrected for control mortality using Abbot's correction

The method for spiking the sediment with the test item was chosen on the basis of the

behaviour ofthe test item in seawater. See pages 10-11 of this report.

UMAA (upper moving average angle) value is calculated manually from the assumption that

the next concentration in the series, i.e. 50000 mg Dodicor V 5654/ kg would give full

mortality.

Prefened pH range is between 7,5 and 8.5, which is natural for seawater. The highest test

concentration (16654 mg Dodicor V 5654/ kg) gave slightly lower pH-values than the

prefen-ed range at day 10.

The test results presented in this report were obtained from tests performed on a sample of

the test item sent to the test facility of Clariant Oil Services, formulated according to the

specifications supplied by the manufacturer or supplier at the time of testing. The test

results cannot be assumed to be valid for samples resulting from subsequent modifications

to the original specifications as supplied.

Description of key information

Two studies with Corophium volutar are available.

In the Key study, based on the nominal concentration of the test item Reaction products of DL-methionine and C18 unsaturated fatty acid chloride and isopropanol, the LC50 for mortality of Corophium volutator PALLAS was > 796 mg/kg sediment dry weight. The NOEC after 10 days was ≥ 796 mg/kg sediment dry weight.

In a supporting study an assessment ofthe solid-phase (sediment) toxicity (lO-day LCso) of Dodicor V 5654 was made

using the sediment-dwelling amphipod, Corophium volutator (Pallas). The following result, expressed

as mg Dodicor V 5654 per kg dry sediment weight, was obtained:

10-day LC50: 7461 mgkg-1 (95% c.i. 3288 - 14821)

Key value for chemical safety assessment

EC10, LC10 or NOEC for marine water sediment:

796 mg/kg sediment dw