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Ecotoxicological information

Long-term toxicity to aquatic invertebrates

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Reference
Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Experimental start date: 14 December 2020 Experimental completion date: 15 July 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
(2012)
Deviations:
yes
Remarks:
For details on deviations please refer to Deviations from Study Plan entry in "Any other information on materials and methods incl. tables" section.
Qualifier:
according to guideline
Guideline:
EU Method C.20 (Daphnia magna Reproduction Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Identification: ALKENES, C15-18 ALPHA-, SULFURIZED
CAS Number: 67762-55-4
EC Number: 267-023-8
Batch: 0720HI434
Purity: 100% UVCB
Physical state / Appearance: Dark red/brown liquid
Expiry date: 01 July 2022
Storage conditions: Room temperature in the dark
Analytical monitoring:
yes
Details on sampling:
Range-finder and Limit Test

The samples were either analyzed on the day of sampling or stored frozen prior to analysis.

Definitive test

The concentration and stability of the test item in the test preparation was verified by chemical analysis on Days 0, 6, 13 and 20 (fresh media) and on Days 1, 7, 14 and 21 (old media)
Vehicle:
yes
Details on test solutions:
Nominal amounts of test item (20, 36, 64, 116 and 200 mg) were each separately added to the surface of 2 L of test water to give the 10, 18, 32, 58 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1 hour. Microscopic inspection of the WAFs showed no micro dispersions or undissolved test item to be present. Photographic images were taken weekly of the control and 100 mg/L loading rate WAF (Fresh) when being viewed under the microscope (see Appendix 14 in attachment “Appendix 1-16”). The aqueous phase or WAF was removed by mid depth siphoning (the first approximate 75 to 100 mL discarded) to give the 10, 18, 32, 58 and 100 mg/L loading rate WAFs.
Test organisms (species):
Daphnia magna
Details on test organisms:
The test was carried out using 1st instar Daphnia magna derived from in-house laboratory cultures.

Adult Daphnia were maintained in 150 mL glass vessels containing Elendt M4 medium (see "Details on Test Conditions" section) in a temperature controlled room at approximately 20 °C. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods. Each culture was fed daily with a mixture of algal suspension (Raphidocelis subcapitata (formally known as Pseudokirchneriella subcapitata)) and GEMMA Micro 300 fish food suspension. Culture conditions ensured that reproduction was by parthenogenesis. Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Each daphnid received approximately 5 to 20 µL of an algal suspension (Raphidocelis subcapitata) daily, supplemented by the addition of approximately 20 µL of GEMMA Micro 300 fish food suspension on Days 0, 1, 2 and 3. Feeding was at a level of approximately 0.1 to 0.2 mg carbon/daphnid/day, dependent on the age and size of the animals. Equal amounts of food were given to each daphnid.
Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
21 d
Hardness:
Water Hardness: 2.5 mmol/L (= 250 mg/L as CaCO3)
Test temperature:
The test vessels were maintained in a temperature controlled room at 18 to 22 °C with a maximum deviation of ± 1 °C
pH:
The pH of the prepared media will be in the range of 7.9 ± 0.3, a pH range of 7.5 to 8.3 (with 0.8 variation) was achieved in control group
Dissolved oxygen:
Dissolved oxygen concentrations were recorded before and after each test media renewal and were measured using a Hach HQ30d Flexi handheld meter.

Dissolved oxygen concentration > 3 mg O2/L throughout the test
Nominal and measured concentrations:
Based on the results of a preliminary range finding test and the initial limit test, Daphnia magna were exposed (10 replicates of a single daphnid per group) to a WAF of the test item over a range of test concentrations of 10, 18, 32, 58 and 100 mg/L loading rates for a period of 21 days. The test solutions were renewed daily throughout the test.

The concentration and stability of the test item in the test preparation was verified by chemical analysis on Days 0, 6, 13 and 20 (fresh media) and on Days 1, 7, 14 and 21 (old media).
Details on test conditions:
For each concentration a single daphnid was placed in 100 mL of the test preparation in 150 mL glass vessels which were then covered with a plastic lid to reduce evaporation. For each test and control group 10 replicate test vessels were prepared. The test vessels were maintained in a temperature controlled room at 18 to 22 °C with a maximum deviation of ± 1 °C with a photoperiod of 16 hours light (not exceeding 1500 Lux) and 8 hours darkness with 20 minute dawn and dusk transition periods for 21 days. The test vessels were not aerated. The diluent water only was aerated prior to use.

The control group was maintained under identical conditions but not exposed to the test item.

The test preparations were renewed daily throughout the test. The adult Daphnia were transferred to fresh media by wide bore pipette before the contents of each vessel were passed through a fine mesh. Young daphnids (live and dead) and any unhatched eggs were collected on the mesh and counted before being discarded.

Each daphnid received approximately 5 to 20 µL of an algal suspension (Raphidocelis subcapitata) daily, supplemented by the addition of approximately 20 µL of GEMMA Micro 300 fish food suspension on Days 0, 1, 2 and 3. Feeding was at a level of approximately 0.1 to 0.2 mg carbon/daphnid/day, dependent on the age and size of the animals. Equal amounts of food were given to each daphnid.

Water Quality Criteria

Dissolved oxygen concentrations, pH and temperature were recorded before and after each test media renewal. The pH and dissolved oxygen concentration were measured using a Hach HQ30d Flexi handheld meter whilst the temperature was measured using a Fisherbrand digital thermometer. Measurements were made on one replicate for each test concentration. The temperature was also measured every hour in a surrogate vessel using a Testo temperature logger. The water hardness of the control and the highest surviving test concentration in the fresh and old media was measured once per week.

The light intensity was also recorded daily throughout the test and was measured by a RS ILM 1332A Light meter.
Test Water

The reconstituted water (Elendt M4 medium) used for the range finding and definitive tests was the same as that used to maintain the stock animals and is defined below:

Reconstituted Water – Elendt M4 Medium

Final concentration in medium

CaCl2.2H2O: 293.8 mg/L
MgSO4.7 H2O: 123.3 mg/L
NaHCO3: 64.8 mg/L
KCl: 5.8 mg/L
Na2SiO3.9 H2O: 10.0 mg/L
NaNO3: 0.274 mg/L
K2HPO4: 0.184 mg/L
KH2PO4: 0.143 mg/L
H3BO3: 2.8595 mg/L
Na2EDTA.2 H2O: 2.50 mg/L
FeSO4.7 H2O: 0.9955 mg/L
MnCl2.4H2O: 0.3605 mg/L
LiCl: 0.306 mg/L
SrCl2.6 H2O: 0.152 mg/L
RbCl: 0.071 mg/L
Na2MoO4.2 H2O: 0.0615 mg/L
CuCl2.2 H2O: 0.0165 mg/L
NaBr: 0.0160 mg/L
ZnCl2: 0.0130 mg/L
CoCl2.6 H2O: 0.010 mg/L
KI: 0.00325 mg/L
Na2SeO3: 0.00219 mg/L
NH4VO3: 0.000575 mg/L
Thiamine hydrochloride: 0.075 µg/L
Cyanocobalamine (Vitamin B12): 0.0010 µg/L
D(+) biotin (Vitamin H): 0.00075 µg/L

Water Hardness: 2.5 mmol/L (= 250 mg/L as CaCO3)
Alkalinity: 0.9 mmol/L

The pH of the prepared media will be in the range of 7.9 ± 0.3 and will be stored at approximately 21 ºC. If necessary the pH may be adjusted with NaOH or HCl.
Reference substance (positive control):
no
Key result
Duration:
21 d
Dose descriptor:
NOELR
Effect conc.:
ca. 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
immobilisation
Key result
Duration:
21 d
Dose descriptor:
NOELR
Effect conc.:
ca. 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
NOELR
Effect conc.:
ca. 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
other: Body Length
Details on results:
For full details on results please refer to "Any other information on results incl. tables" section.

For numbered tables mentioned below please refer to relevant attachment.

For numbered Appendixes mentioned below please refer to attachment “Appendix 1-16”.

Range-finding Test

Cumulative immobilization and sub lethal effects data from the exposure of Daphnia magna to the test item during the range finding test are given in Table 1.

No immobilization was observed at the test concentrations of 1.0, 10 and 100 mg/L loading rate WAF’s. Sub lethal effects of exposure were observed at a test concentration of 10 mg/L loading rate WAF. This response was noted in one parental Daphnid only, which appeared ‘small and pale’.

Chemical analysis of the test preparations on Days 0 and 1 showed measured test concentrations to range from 0.039 to 3.8 mg/L. Chemical analysis of the expired test preparations on Day 1 could not detect any test item (the limit of detection was determined to be 0.010 mg/L) indicating that the test item was unstable under test conditions.

Initial Limit Test

No immobilization or sub lethal effects of exposure were observed at the test concentration of 100 mg/L loading rate WAF. However, the control group exhibited 10% mortality throughout the test with sub lethal effects of exposure of the daphnids appearing pale and covered in debris.

Analysis of the fresh test preparations on Days 0, 6, 13 and 20 showed measured test concentrations to range from 3.3 to 6.4 mg/L. A decline in measured test concentration of the aged test preparations on Days 1, 7, 14 and 21 was observed to be between less than the limit of quantification (LOQ) which was determined to be 0.11 mg/L and 0.65 mg/L.

Definitive Test

Based on the results of a preliminary range finding test and initial limit test, Daphnia magna were exposed (10 replicates of a single daphnid per group) to a WAF of the test item over a range of test concentrations of 10, 18, 32, 58 and 100 mg/L loading rates for a period of 21 days. The test solutions were renewed daily throughout the test and were analyzed for concentration as a more sensitive method of analysis was required in order to determine test concentrations at the lower concentration being utilized during the definitive test.

Verification of Test Concentrations

Analysis of the fresh test preparations on Days 0 , 6*, 13 and 20 showed measured test concentrations to range from 0.021 to 1.1 mg/L. A decline in measured test concentration of the aged test preparations on Days 1*, 7*, 14 and 21 was observed to be between less than the limit of quantification (LOQ) which was determined to be 0.005 mg/L and 0.41 mg/L.  

The dissolved fraction of the WAF test solution may have been comprised of one or several components of the test item. Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

*Days 0 and 1 were based on the means of the original and duplicate test samples, Days 6 and 7 were based on the duplicate test samples only as procedural recoveries failed during the original sample analysis.

Test Organism Observations

The observations for each test and control group are summarized in Table 3. The total cumulative production of live young is given in Table 4 and the number of live young produced per adult is shown in Table 2. The total number of offspring per parent Daphnia (for each replicate) alive at the end of the test is shown in Figure 1 (please refer to attachment “Figure 1”). Data for each individual replicate vessel are given in Appendix 2 to Appendix 7.

Validation Criteria

The following validation criteria were achieved during the test:

 

Required

Actual

Control mortality

≤ 20%

20%

Mean number of live young per surviving adult (control group)

≥ 60 after 21 days

129

Coefficient of variation for control group†

≤ 25%

14.2%

No ephippia produced

0

0

Dissolved oxygen

> 3 mg O2/L

≥ 8.3 mg O2/L

pH (control group)

6 to 9
variation ≤1.5

7.5 to 8.3
0.8

† Based on total number of living offspring per parent animal alive at the end of the test

Lethal Effects on the Parental Generation (P1)

The control group exhibited 20% mortality throughout the test whereas lower levels of mortality (10%) were observed at the test loading rates of 32, 58 and 100 mg/L. Statistical analysis of the mortality data using Fisher’s Exact Binomial Test with Bonferroni Correction Test Procedure showed that the observed mortalities in the 32, 58 and 10 mg/L loading rate test groups were not significantly different (P≥0.05) when compared to the control. No mortalities occurred in the 10 and 18 mg/L loading rate test groups throughout the test (see Appendix 1).

The following ELx, NOEL and LOEL (immobilization) values based on nominal loading rates were estimated by inspection of the data or calculated by Fisher’s Exact Binomial Test with Bonferroni Correction Test Procedure incorporating Qualitative Trend analysis by Contrasts at 21 days:

Endpoint

Nominal Loading Rate mg/L

(95% confidence limits)*

Immobilization

EL10

>100 (not determined)

EL20

>100 (not determined)

EL50

>100 (not determined)

No Observed Effect Loading Rate

100

Lowest Observed Effect Loading Rate

Not determined

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

* It was not possible to calculate 95% confidence limits for the ELx values as the data generated did not fit the models available for the calculation of confidence limits.

Sub-lethal Effects on the Parental Generation (P1)

Sub lethal effects of exposure were observed (including prior to one Daphnia being found dead in the control and at 58 and 100 mg/L loading rates) in the control and at all test concentrations; observations included reduced mobility, small, pale and trapped at surface. These observations generally persisted from Days 5 to 11. However, in the control group one daphnid was observed to have reduced mobility on Day 17 and was observed to be pale with reduced mobility from Days 18 to 20 and at the 32 mg/L loading rate one daphnid was observed to be pale on Day 18.

After 21 days the length of each surviving adult was determined, the results of which are given in Table 5. The results showed that no significant differences (P≥0.05) were found between the control and each of the test groups in terms of length of the surviving parental Daphnids on Day 21 using the below methods of statistical analysis (see Appendix 1).

The following ELx, NOEC and LOEC values (body length) based on geometric mean measured test concentrations were calculated using the 3-Parameter Normal Cumulative Distribution Function and the Dunnett’s Multiple t-test Procedure incorporating Shapiro Wilk's test on normal distribution, Levene’s test on variance homogeneity and Trend analysis by Contrasts at 21 days:

Endpoint

Nominal Loading Rate mg/L

(95% confidence limits)*

Body Length

EL10

>100 (not determined)

EL20

>100 (not determined)

EL50

>100 (not determined)

No Observed Effect Loading Rate

100

Lowest Observed Effect Loading Rate

Not determined

* It was not possible to calculate 95% confidence limits for the ELx values as the data generated did not fit the models available for the calculation of confidence limits.

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

Effects on Reproduction

After 21 days there were no statistically significant differences (P≥0.05) between the control and any of the loading rate test groups (see Appendix 1).

The following ELx, NOEL and LOEL (reproduction) values based on geometric mean measured test concentrations were calculated by Probit analysis using Linear-maximum likelihood regression and Williams Multiple Sequential t test Procedure incorporating Shapiro Wilk's test on normal distribution, Levene’s test on variance homogeneity and Trend analysis by Contrasts at 21 days:

Endpoint

Nominal Loading Rate mg/L

(95% confidence limits)*

Reproduction

EL10

>100 (not determined)

EL20

>100 (not determined)

EL50

>100 (not determined)

No Observed Effect Loading Rate

100

Lowest Observed Effect Loading Rate

Not determined

* It was not possible to calculate 95% confidence limits for the ELx values as the data generated did not fit the models available for the calculation of confidence limits.

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

Effects on the Filial Generation (F1)

Information on the effects of the test item on the F1 generation is limited, since, by study design, the young are removed soon after liberation from the brood pouch. However, an assessment was made at each media renewal with data for each individual replicate vessel are given in Appendix 8 to Appendix 13.

Young were first produced in the control test group on Day 8 of the test.

Numbers of unhatched eggs and dead young were low in all control and treatment groups surviving to maturation.

Lowest Observed Effect Loading Rate

The LOEL could not be determined, as at the highest loading rate tested (100 mg/L) no significant mortalities (immobilization) were observed in the parental generation (P1), there were no significant differences (P≥0.05) between the control and the 100 mg/L loading rate WAF group in terms of numbers of live young produced per adult by Day 21 and there were no significant differences (P≥0.05) in terms of the average adult body length when compared to the control after 21 days.

No Observed Effect Loading Rate

The NOEL was 100 mg/L loading rate WAF as there were no significant mortalities (immobilization) were observed in the parental generation (P1), there were no significant differences (P≥0.05) between the control and the 100 mg/L loading rate WAF group in terms of numbers of live young produced per adult by Day 21 and there were no significant differences (P≥0.05) in terms of the average adult body length when compared to the control after 21 days.

Water Quality Criteria

The results of the water quality measurements are given in Table 6.

Temperature was maintained at approximately 18 °C to 22 °C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

The results of the water hardness measurements are given in Table 7.

The water hardness was observed to be in the range 246 to 280 mg/L as CaCO3 in the control and the highest surviving test group throughout the test.

Throughout the test the light intensity was observed to be in the range 747 to 998 lux (see Table 8).

The water temperature was also recorded in the control vessel every hour using a Testo temperature logger (see Figure 2, please refer to attachment “Figure 2”) which was maintained at approximately 20 °C to 21 °C.

Vortex Depth Measurements

The vortex depth was recorded at the start and end of the mixing period and was observed to have formed a dimple at the media surface.

Observations on Test Item Solubility

Observations on the test media were carried out during the mixing and testing of the WAFs.

At the start of the mixing period the 10, 18, 32, 58 and 100 mg/L loading rates were observed to be a clear colorless water column with test item floating at the surface. After 23 hours stirring and a 1 Hour standing period the 10, 18, 32, 58 and 100 mg/L loading rates remained as at the start of stirring. Microscopic inspection of the WAF showed no micro dispersions or undissolved test item to be present. After siphoning and for the duration of the test, the 10, 18, 32, 58 and 100 mg/L loading rates were observed to be clear, colorless solutions.

At the start and throughout the test all control solutions were observed to be clear colorless solutions.

Photographic Image of Treatment Groups

For illustrative purposes only, collective photographic images of the control and 100 mg/L loading rate WAF treatment groups are given in Appendix 14.

The images illustrate that the 100 mg/L loading rate WAF preparation did not contain any undissolved test item and as such the images at 100 mg/L loading rate were comparable to the control.

Validity criteria fulfilled:
yes
Conclusions:
It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

For tabulated endpoints please refer to table in "Executive Summary".
Executive summary:

Introduction

A study was performed to assess the chronic toxicity of the test item to Daphnia magna.  The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (2012) No 211, "Daphnia magna Reproduction Test" referenced as Method C.20 of Commission Regulation (EC) No. 440/2008.

Methods

Due to the low aqueous solubility and complex nature of the test item, the test medium was prepared as a Water Accommodated Fraction (WAF).

Based on the results of a preliminary range‑finding test and an initial limit test, Daphnia magna were exposed (10 replicates of a single daphnid per group) to a WAF of the test item over a range of nominal loading rates of 10, 18, 32, 58 and 100 mg/L for a period of 21 days.  Nominal amounts of test item (20, 36, 64, 116 and 200 mg) were each separately added to the surface of 2 L of test water to give the 10, 18, 32, 58 and 100 mg/L loading rates respectively.  After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface.  The stirring was stopped after 23 hours and the mixture allowed to stand for 1 hour.  The aqueous phase or WAF was removed by mid depth siphoning (the first approximate 75-100 mL discarded) to give the 10, 18, 32, 58 and 100 mg/L loading rate WAFs.  Microscopic inspection of the WAF showed no micro dispersions or undissolved test item to be present.

The test solutions were renewed daily throughout the test. 

The numbers of live and dead adult and young daphnids were determined daily.  The daphnids were fed daily with a mixture of algal suspension and GEMMA Micro 300 food suspension.

Results

Analysis of the fresh test preparations on Days 0 , 6*, 13 and 20 showed measured test concentrations to range from 0.021 to 1.1 mg/L. A decline in measured test concentration of the aged test preparations on Days 1*, 7*, 14 and 21 was observed to be between less than the limit of quantification (LOQ) which was determined to be 0.005 mg/L, and 0.41 mg/L.

The dissolved fraction of the WAF test solution may have been comprised of one or several components of the test item. Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only, as recommended in OECD guidance document 23 (2019).

* Days 0 and 1 were based on the means of the original and duplicate test samples, Days 6 and 7 were based on the duplicate test samples only as procedural recoveries failed during the original sample analysis.

Exposure of Daphnia magna to the test item gave the following results based on nominal loading rates:

Endpoint

Nominal Loading Rate
mg/L
(95% confidence limits)[1]

Immobilization

EL10

>100 (not determined)

EL20

>100 (not determined)

EL50

>100 (not determined)

No Observed Effect Loading Rate (NOEL)

100

Lowest Observed Effect Loading Rate (LOEL)

Not determined

Reproduction

EL10

>100 (not determined)

EL20

>100 (not determined)

EL50

>100 (not determined)

No Observed Effect Loading Rate (NOEL)

100

Lowest Observed Effect Loading Rate (LOEL)

Not determined

Body Length

EL10

>100 (not determined)

EL20

>100 (not determined)

EL50

>100 (not determined)

No Observed Effect Loading Rate (NOEL)

100

Lowest Observed Effect Loading Rate (LOEL)

Not determined

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

 

[1]        It was not possible to calculate 95% confidence limits for the ELx values as the data generated did not fit the models available for the calculation of confidence limits.

Description of key information

A study was performed to assess the chronic toxicity of the test item to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (2012) No 211, "Daphnia magna Reproduction Test" referenced as Method C.20 of Commission Regulation (EC) No. 440/2008. Due to the low aqueous solubility and complex nature of the test item, the test medium was prepared as a Water Accommodated Fraction (WAF).

Based on the results of a preliminary rangefinding test and an initial limit test, Daphnia magna were exposed (10 replicates of a single daphnid per group) to a WAF of the test item over a range of nominal loading rates of 10, 18, 32, 58 and 100 mg/L for a period of 21 days. Nominal amounts of test item (20,36, 64, 116 and 200 mg) were each separately added to the surface of 2 L of test water to give the 10, 18, 32, 58 and 100 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface.  The stirring was stopped after 23 hours and the mixture allowed to stand for 1 hour.  The aqueous phase or WAF was removed by mid depth siphoning (the first approximate 75-100 mL discarded) to give the 10, 18, 32, 58 and 100 mg/L loading rate WAFs.  Microscopic inspection of the WAF showed no micro dispersions or undissolved test item to be present. The test solutions were renewed daily throughout the test.

The results were as follows:

Endpoint

Effect level

Nominal loading rate (mg/L)

Immobilisation

EL10

>100

EL20

>100

EL50

>100

NOEL

>100

LOEL

Not determined

Reproduction

EL10

>100

EL20

>100

EL50

>100

NOEL

>100

LOEL

Not determined

Body length

EL10

>100

EL20

>100

EL50

>100

NOEL

>100

LOEL

Not determined

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates
Dose descriptor:
EC10
Effect concentration:
> 100 mg/L

Additional information