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Long-term toxicity to aquatic invertebrates

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Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-03-13 to 2018-11-21
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
requested from ECHA final decision on substance evaluation
Qualifier:
according to
Guideline:
OECD Guideline 211 (Daphnia magna Reproduction Test)
Version / remarks:
2012
Deviations:
no
GLP compliance:
yes (incl. certificate)
Analytical monitoring:
yes
Remarks:
LC-MS/MS
Details on sampling:
- Concentrations: 0.0100 - 0.0316 - 0.100 - 0.316 - 1.00 - 3.16 mg/L corresponding to 0.000460 – 0.00145 – 0.00460 – 0.0145 – 0.0460 – 0.145 mg pure compound/L.
- Sampling method: All concentration levels and the control were analyzed via LC-MS/MS in fresh media at the start of the exposure intervals on days 0, 6, 14 (0 hours) and in old media at the end of the exposure intervals on days 2, 8, 16 (48 hours).
- Sample storage conditions before analysis: All original samples were stored at room temperature. Prepared samples were stored at room temperature in an autosampler until analysis.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): A stock solution (3.16 mg/L of the test item were weighed out) was stock solution freshly prepared with test medium (as specified below) for each exposure interval. The stock solution was stirred with a magnetic stirrer (1100 rpm for 30 minutes at room temperature) and was used for preparation of the test concentrations. 6 concentration levels of the test item in a geometric series with a separation factor of √10, prepared by diluting the stock solution of 3.16 mg/L with test medium (as specified below), were tested as follows: 0.0100 - 0.0316 - 0.100 - 0.316 - 1.00 - 3.16 mg/L corresponding to 0.000460 – 0.00145 – 0.00460 – 0.0145 – 0.0460 – 0.145 mg pure compound/L. The concentrations were based on the results of a preliminary test.

- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Strain/clone: Daphnia magna STRAUS (Clone 5)
- Source: Institut Dr. Nowak, Mayenbrook 1, 28870 Ottersberg, Germany
- Age of parental stock (mean and range, SD): 2 to 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 22 hours before the start of the exposure and discarded. The juveniles born within the following period of max. 22 hours preceding the exposure were used for the test after an acclimatization phase of at least 2 hours in the test medium (as specified above). No first brood progeny was used for the test.
- Feeding during test
- Food type: Pseudokirchneriella subcapitata and Desmodesmus subspicatus suspension was provided as food.
- Amount: max. 0.2 mg C per Daphnia and day.
- Frequency: Daily feeding per test vessel
- Health during acclimation (any mortality observed): not observed
ACCLIMATION
- Acclimation period: The juvenile daphnids were acclimatized in the test medium (natural river water supplemented with mineral components of the culture medium as specified above) for at least 2 hours prior to insertion in the test solutions. An acclimatization of the brood daphnids 3 weeks prior to the test as suggested by the guideline is not necessary, since the test medium is supplemented with the same mineral components as the culture medium.
- Acclimation conditions (same as test or not): yes
- Type and amount of food: Pseudokirchneriella subcapitata and Desmodesmus subspicatus suspension was provided as food.
- Feeding frequency: Daily feeding
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Hardness:
42.7 mg CO3/L
Test temperature:
The temperature was in the range of 18 – 22 °C and should not vary by more than 2°C.
Dissolved oxygen:
9.11 mg O2/L
Conductivity:
79.6 µS/cm
Nominal and measured concentrations:
Nominal: 0.0100 - 0.0316 - 0.100 - 0.316 - 1.00 - 3.16 mg/L corresponding to 0.000460 – 0.00145 – 0.00460 – 0.0145 – 0.0460 – 0.145 mg pure compound/L
The concentrations of the test item were analytically verified via LC-MS/MS in fresh and old media. Four compounds have been detected and quantified as a group. Although the test item concentrations varied to a larger extent nominal concentrations are used to derive effect levels as the test item is an UVCB substance.
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass beakers (5 (ID) x 8 (H) cm), 100 mL capacity, loosely covered with watch glasses
- Type (delete if not applicable): loosely covered with watch glasses
- Material, size, headspace, fill volume: Glass beakers (5 (ID) x 8 (H) cm), 100 mL capacity, Test volume 50 mL
- Aeration: Test vessels were not aerated during the test.
- Renewal rate of test solution (frequency/flow rate): The test solutions were renewed every 48 hours during exposure. For this purpose, a second set of test vessels was filled with the freshly prepared test solutions and the daphnids were transferred by pipette
- No. of organisms per vessel: 10 daphnids in 10 replicates were used for all concentration levels and the control with one daphnid per replicate, which was held individually.
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
Water Parameters of the diluted River Water
River Grane
Location Im Granetal, 38685 Langelsheim, Germany Latitude:51.9146°, Longitude: 10.3819°
Sampling Date 2017-10-26
Weather on Day of Sampling Cloudy; 15°C
Colour Yellowish to brownish, clear
pH 7.90
Conductivity [µS/cm] 79.6
Dissolved oxygen [mg O2/L] 9.11
TOC [mg C/L] 2.46
Ammonium-N [mg NH4-N/L] < 0.04
Nitrate-N [mg NO3-N/L] 0.30
o-Phosphate-p [mg P/L] < 0.20
Total phosphate [mg P/L] < 0.20
Suspended matter [mg/L] 7.00
Total hardness [mg CO3/L] 42.7


- Culture medium different from test medium: yes, Culture medium: Elendt M4, according to OECD 211, Annex 2, Test medium River water supplemented with culture medium

OTHER TEST CONDITIONS
- Photoperiod: 16/8 hours light/dark cycle
- Light intensity: Max. 1500 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality of parental : The number of parental daphnids, which were immobilized or dying daphnids during the test and the dates of death were observed and recorded once a day at least at the same time as the offspring was counted. Dead specimens were removed as soon as they were detected.

Offspring: First appearance of juveniles was checked daily. The number of neonates (alive and dead progeny) was counted and dates of release were recorded once a day from the first day of appearance of neonates until the end of the test. The neonates (alive and dead) were removed after counting and before addition of algae to prevent them from consuming food intended for the adults. The number of aborted eggs or dead offspring and dates of observation were recorded.

Additional observations /Abnormalities (e.g. abnormal swimming behavior, number of males Condition of parental and winter eggs) were observed and recorded on each day of daphnids observation.

Total body length / Mean dry : At the end of the test, the total length excluding the anal spine of weight of parental daphnids each survived parental daphnid and the mean dry weight of the survived parental daphnids of all test concentrations and the control were determined. As stated in the OECD protocol, this may be useful additional information but is not used for determination of a NOEC.

Additional endpoints: The time to first brood, the intrinsic rate of population increase and the number and size of first brood per animal were reported, but not used for endpoint calculations.



TEST CONCENTRATIONS
- Spacing factor for test concentrations: factor of √10
- Range finding study, yes, non GLP
- Test concentrations: Four nominal concentrations of 0.001 – 0.01 – 0.10 and 1.00 mg/L were tested.
Reference substance (positive control):
yes
Remarks:
potassium dichromate
Key result
Duration:
21 d
Dose descriptor:
EC10
Effect conc.:
> 145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
EC50
Effect conc.:
> 145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
> 145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
>= 145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Key result
Duration:
21 d
Dose descriptor:
other: LC20
Effect conc.:
> 145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adults
Key result
Duration:
21 d
Dose descriptor:
LC50
Effect conc.:
> 145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adults
Key result
Duration:
21 d
Dose descriptor:
LOEC
Effect conc.:
145 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adults
Key result
Duration:
21 d
Dose descriptor:
NOEC
Effect conc.:
46 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks:
adults
Details on results:
- Behavioural abnormalities:
- Observations on body length and weight: The mean values of the body length (excluding the anal spine) of the survived parental daphnids in all tested concentration levels were in the range of 4.60 to 4.73 mm per daphnid and 4.78 mm per daphnid in the control group.
The mean dry weight of the survived parental daphnids was in the range of 0.850 to 0.970 mg per daphnid in all tested concentration levels and 1.02 mg per daphnid in the control

- Other biological observations: The first appearance of living juveniles was observed between days 8 and 12 at all survived parental daphnids of the control and all tested concentration levels. Stillborn juveniles and aborted eggs were observed during the exposure period of 21 days. No males were observed in the control or in the test groups during the test. No ephippia were observed in the control or in the test groups during the test.
- Mortality of control: Accidental and inadvertent mortality as defined in the OECD test guideline were not observed.
Adult mortality was not observed in the control and the nominal pure compound concentrations of 0.460, 1.45, 4.60 and 46.0 µg/L. At the pure compound concentration 14.5 and 145 µg/L, mortalities of 10 and 20 % of the introduced adults was observed, respectively. A statistical significant difference was found for this parameter at the highest concentration of 145 µg/L, when tested with the step-down Cochran-Armitage test procedure (α = 0.05).
Results with reference substance (positive control):
- Results with reference substance valid? yes, The percentage of immobility for the reference item potassium dichromate (SIGMA-ALDRICH, batch number MKBV0900V, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours from 2018-03-13 to 2018-03-14.
- Relevant effect levels: EC50 = 1.66 mg/L, Confidence limits = 1.00 - 4.00 mg/L, Valid Range: 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5; 0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A

Reported statistics and error estimates:
The NOEC and LOEC for the reproduction and the adult mortality were determined with the software ToxRat Professional as specified below.
Significant deviations of reproduction were determined in comparison to the control using statistical standard procedures as normality test (Shapiro-Wilk’s test), variance homogeneity test (Levene’s test) and a one-sided smaller multiple sequentially-rejective median test after Bonferroni-Holm (for reproduction).
Prior to running the multiple test, a normality test, a variance homogeneity test and a trend analysis were performed. P values of the normality and the variance homogeneity test were 0.01 and 0.05 for the trend analysis. The α-value (acceptable probability of incorrectly concluding that there is a difference) was α = 0.01 for the normality test and the variance homogeneity test and α = 0.05 for the trend analysis. For the effects on reproduction, the juveniles per introduced parent at the start of the exposure excluding accidental and inadvertent mortality (reproductive output) were calculated. Since no accidental and inadvertent mortality was observed, no parental daphnids were excluded from the evaluation of the reproductive output. The effects on reproduction per survived parental animal at the end of the test were also calculated. For the determination of significant deviations of the adult mortality, step-down Cochran-Armitage test procedure was used. The coefficients of variation around the mean number of living offspring produced per introduced parent and per survived parent in the control and the test groups were evaluated.
All effect levels (NOEC, LOEC, EC10 / 50 / 100 and LC20 / 50 / 100) given are based on nominal concentrations of the pure compound of the test item. The EC10 / 50-values for the reproductive output were empirically EC-/LC-values derived from the observed data, since no significant reduction was observed.

Effects on Reproduction for all Introduced and for Surviving Parents

Nominal concentration of the pure compound of the test item

Mean number of offspring
per introduced parental daphnid

Mean number of offspring
per survived parental daphnid

[µg/L]

Mean

SD

CV

Mean

SD

CV

145

120.4

67.7

56.2

150.5

26.6

17.7

  46.0

118.6

34.1

28.8

118.6

34.1

28.8

  14.5

117.5

40.9

34.8

129.0

19.8

15.3

    4.60

137.5

18.0

13.1

137.5

18.0

13.1

    1.45

119.3

33.8

28.3

119.3

33.8

28.3

    0.460

115.6

26.7

23.1

115.6

26.7

23.1

Control

129.5

21.2

16.4

129.5

21.2

16.4

Mortality [%] of the Adult Daphnids after 7, 14 and 21 Days of Exposure

Nominal concentration
of the pure compound
of the test item

[µg/L]

Adult Mortality [%]

7 days

14 days

21 days

145

20

20

 201)

  46.0

0

0

0

  14.5

0

10

10

    4.60

0

0

0

    1.45

0

0

0

    0.460

0

0

0

Control

0

0

0

First Appearance of Living Juveniles in the Individual Groups

Nominal concentration
of the pure compound
of the test item

[µg/L]

Day of first appearance of living juveniles at the survived parental daphnids in replicate no.

First

appearance

1

2

3

4

5

6

7

8

9

10

mean day

145

--

8

8

10

8

8

--

9

9

8

8.5

     46.0

9

9

9

9

9

8

9

10

8

8

8.8

     14.5

8

8

10

8

8

10

10

8

9

9

8.8

         4.60

8

9

10

9

9

8

9

9

9

10

9.0

         1.45

10

8

9

9

9

8

9

9

9

8

8.8

           0.460

8

11

9

8

9

9

9

8

12

8

9.1

Control

8

9

11

9

8

9

9

11

8

8

9.0

Total Body Length and Dry Weight of the Survived Parental Daphnids

Nominal concentration
of the pure compound
of the test item

[µg/L]

Total length of the survived parent animals [mm]

MV

N

Dry weight

[mg]

Replicate no.

 

 

1

2

3

4

5

6

7

8

9

10

[mm]

 

S

MV

145

--

4.25

4.75

4.75

4.75

4.50

--

4.75

4.75

4.75

4.66

8

6.80

0.850

     46.0

4.50

4.50

5.00

4.50

4.50

4.50

4.75

4.50

4.50

5.00

4.63

10

9.00

0.900

     14.5

5.00

4.50

4.50

4.75

5.00

--

5.00

4.75

4.50

4.50

4.72

9

8.40

0.930

         4.60

4.75

4.75

4.50

4.50

4.75

4.75

4.75

4.50

4.75

4.75

4.68

10

9.70

0.970

         1.45

4.50

4.75

4.50

4.75

4.75

4.50

4.50

4.50

4.50

4.75

4.60

10

8.70

0.870

           0.460

5.00

4.75

4.50

4.75

4.75

4.50

4.75

4.75

4.50

5.00

4.73

10

8.70

0.870

Control

5.00

4.50

4.50

4.75

5.00

5.00

4.50

4.75

5.00

4.75

4.78

10

10.2

1.02

Validity criteria fulfilled:
yes
Conclusions:
The effects on reproduction were evaluated based on the reproduction per introduced animal, since the ecologically most relevant response variable is the total number of living offspring produced per parent animal which does not die accidentally or inadvertently during the test.
The overall effect threshold for effects of the test item under the test conditions was >145 µg/L (EC20 adults mortality, EC10 reproduction) 46.0 µg/L (NOEC adult mortality) and 145 µg/L (LOEC reproduction and LOEC adults mortality) based on the nominal concentrations of the pure compound of the test item HH-2014-548.
Executive summary:

The 21-day-chronic toxicity of HH-2014-548 to Daphnia magna was studied under semi-static conditions. Test species were exposed to control and test chemical at nominal concentrations of 0.000460 – 0.00145 – 0.00460 – 0.0145 – 0.0460 – 0.145 mg pure compound/L.  The 21-day EC10 reproduction/EC20 adult mortalitywas > 145 µg a.i./L.  The 21-day NOEC based on adult mortality was 46 µg a.i./L.  The effect occured was an insignificant increase of mortality (10%) at a concentration of 14.5 µg/L. However, this effects was not significant and there was no dose dependent increase in mortality, thus, this effect was considered not toxicologically relevant. Production of offspring in the treated groups indicated that HH-2014-548 did not have an effect on the reproduction at concentrations greater than 145 µg a.i./L.  The most sensitive end point was total number of living offspring produced per parent animal.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1stinstar):

Test Type: Semi-static

 

EC20: >145 µg a.i./L

NOEL:  46 µg a.i./L

LOEC:  145 µg a.i./L}                     Endpoint(s) Effected:  adult mortality

 

Endpoint:
long-term toxicity to aquatic invertebrates
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: scientifically acceptable study, that satisfies basic scientific requirements, however not all data are reported
Guideline:
other: Coupled OECD Confirmatory test
GLP compliance:
not specified
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Coupled OECD Confirmatory test, Degradation products from an activated sludge unit
- Effluent: 0.2 mg/L ammonia
- Controls: activated sluge unit without test mat. was run in parallel
Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Water flea
- Age of parental stock (mean and range, SD): <= 24 h
- Feeding during test: yes
- Food type: green algae: Scenedesmus subspicatus (New name: Desmodesmus subspicatus)
- Amount: feed was calculated to be sufficient for 24 h
- Frequency: daily

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
- new borne daphnids were removed and counted daily
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
21 d
Remarks on exposure duration:
per generation
Details on test conditions:
TEST SYSTEM
- Test vessel: 500mL glass beaker
- Type: open
- Aeration: No
- Renewal rate of test solution (frequency): daily diluted bulk sample of the effluent of the model activated sluge unit, dilution was 1:1.25
- No. of organisms per vessel: 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The influent of the activated sludge unit consisted of synthetic wastewater with a 10 mg/L concentration of REWOQUAT W 90. A control activated sludge unit without test item was run in parallel.
- dilution water: filtered drinking water (charcoal)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- reproduction rate after 21 days, growth rate, parental mortality

Reference substance (positive control):
no
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
> 100 µg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
reproduction
Validity criteria fulfilled:
yes
Remarks:
Validity criterium was fulfilled: reproduction rate was 40 - 60 neonates
Conclusions:
The 21-day-reproduction test of the effluent of a model activated sludge unit fed with 10 mg/L partially unsaturated IQAC, DMS quaternised to Daphnia magna was studied under static renewal conditions. Daphnids were exposed to the effluent of the activated sludge unit containing the degradation products of the test item. A control activated sludge unit without test item was run in parallel. During the test partially unsaturated IQAC, DMS quaternised, is eliminated in the model activated sludge unit by > 90%. The elimination is calculated to be 97.75% taking into account the inflow into the model activated sludge unit of the test substance with a concentration of 10 mg/l, the dilution of the effluent of the elimination unit in ratio 1:1.25 and the separately determined EC0=0.1 mg/l. Thus a NOEC of >100 μg/L is calculated.
Executive summary:

The 21-day-reproduction test of the effluent of a model activated sludge unit fed with 10 mg/L partially unsaturated IQAC, DMS quaternised to Daphnia magna was studied under static renewal conditions. Daphnidswere exposed to the effluent of the activated sludge unit containing the degradation products of test item. A control activated sludge unit without test item was run in parallel. The test duration for every generation was 21 days. Neonates of the 14thtest day were exposed as filial generation under the same conditions as the parental generation. Filial generations 1 and 2 were monitored for reproduction rate, time of the first offspring and parental mortality.

Parental generation:

In the parental generation there was no difference between the exposed and the not exposed daphnids, however the first offspring in the controls was one day later. Mortality of the parental generation was slightly higher in the control.

F1 generation:

42 days after the start of exposition of parental generation, the reproduction rateof the surfactant exposed F1 generation was enhanced and exeeded that of the control. The mean number of parental daphnids was greater than in the control. There was no difference in the time of appearance of the first offspring. Mortality of the parental generation was slightly higher in the control.

F2 generation:

63 days after the start of exposition of the first parental generation, in the secondfilial generation some adverse effects were observed for the surfactant exposed Daphnids: Growth rate decreased and the size and colour of the control animals were not reached. The reproduction rate was significant lower than in the parental generation and the F1 generation. The mortality of the parental animals increased .

During the test partially unsaturated IQAC, DMS quaternised, is eliminated in the model activated sludge unit by > 90%. The elimination is calculated to be 97.75% taking into account the inflow into the model activated sludge unit of the test substance with a concentration of 10 mg/l, the dilution of the effluent of the elimination unit in ratio 1:1.25 and the separately determined EC0=0.1 mg/l. Thus a NOEC of >100 μg/L is calculated.

The test material contains isopropanol. The toxicity of isopropanol against Daphnia magna is more than 10 000 times lower than the toxicity of the test material. Water solubility of the test item may be slightly enhanced leading to a better bioavailability and slightly enhanced toxicity. But these effects are regarded to have a minor influence on the test result.Therefore it can be concluded in a first approach, neglecting additive effects, that the observed effect values can be attributed to the active ingredient itself.

This study is classified as acceptable and satisfies basic scientifical requirements for a chronic toxicity study with freshwater invertebrates, allthough not all data were reported.

Results Synopsis
Test Organism Age (1stinstar): Daphnia magna
Test Type: Static Renewal 63d
NOEC : >100 μg/L (calculated)
Endpoints Effected:  reproduction rate, time of the first offspring, growth and parental mortality

Description of key information

The 21-day-chronic toxicity of Oleic acid based IQACs, DMS quaternised to Daphnia magna was studied under semi-static conditions. Test species were exposed to control and test chemical at nominal concentrations of 0.000460 – 0.00145 – 0.00460 – 0.0145 – 0.0460 – 0.145 mg pure compound/L. The overall effect threshold for effects of the test item under the test conditions was >145 µg/L (EC20 adults mortality, EC10 reproduction) 46.0 µg/L (NOEC adult mortality) and 145 µg/L (LOEC reproduction and LOEC adults mortality) based on the nominal concentrations of the pure compound of the test item HH-2014-548.

 

Additionally, the 63d NOEC of >100 μg/L was determined in a 21-day-reproduction test with Daphnia magna over 3 generations. Daphnids were exposed to the effluent of the activated sludge unit containing the degradation products of partially unsaturated IQAC, DMS quaternised.

Key value for chemical safety assessment

EC10, LC10 or NOEC for freshwater invertebrates:
145 µg/L

Additional information

The 21-day-chronic toxicity of Oleic acid based IQACs, DMS quaternised to Daphnia magna was studied under semi-static conditions. Test species were exposed to control and test chemical at nominal concentrations of 0.000460 – 0.00145 – 0.00460 – 0.0145 – 0.0460 – 0.145 mg pure compound/L.  The 21-day EC10 reproduction/EC20 adult mortalitywas > 145 µg a.i./L.  The 21-day NOEC based on adult mortality was 46 µg a.i./L.  The effect occured was an insignificant increase of mortality (10%) at a concentration of 14.5 µg/L. However, this effects was not significant and there was no dose dependent increase in mortality, thus, this effect was considered not toxicologically relevant. Production of offspring in the treated groups indicated that HH-2014-548 did not have an effect on the reproduction at concentrations greater than 145 µg a.i./L.  The most sensitive end point was total number of living offspring produced per parent animal.

 

This study is classified as acceptable and satisfies the guideline requirements for a chronic toxicity study with freshwater invertebrates.

 

Results Synopsis

 

Test Organism Age (e.g. 1stinstar):

Test Type: Semi-static

 

EC20: >145 µg a.i./L

NOEL:  46 µg a.i./L

LOEC:  145 µg a.i./L}                     Endpoint(s) Effected:  adult mortality

The 21-day-reproduction test of the effluent of a model activated sludge unit fed with 10 mg/Lpartially unsaturatedIQAC (containing 10% of Isopropanol) to Daphnia magna was studied under static renewal conditions.Daphnidswere exposed to the effluent of the activated sludge unit containing the degradation products of the test item. A control activated sludge unit without the test item was run in parallel. The test duration for every generation was 21 days. Neonates of the 14thtest day were exposed as filial generation under the same conditions as the parental generation. Filial generations 1 and 2 were monitored for reproduction rate, time of the first offspring and parental mortality.

Parental generation:

In the parental generation there was no difference between the exposed and the not exposed daphnids, however the first offspring in the controls was one day later. Mortality of the parental generation was slightly higher in the control.

F1 generation:

42 days after the start of exposition of parental generation, the reproduction rate of the surfactant exposed F1 generation was enhanced and exceeded that of the control. The mean number of parental daphnids was greater than in the control. There was no difference in the time of appearance of the first offspring. Mortality of the parental generation was slightly higher in the control.

F2 generation:

63 days after the start of exposition of the first parental generation, in the second filial generation some adverse effects were observed for the surfactant exposed Daphnids: Growth rate decreased and the size and colour of the control animals were not reached. The reproduction rate was significant lower than in the parental generation and the F1 generation. The mortality of the parental animals increased.

During the test the read across substance, partially unsaturated IQAC, DMS quaternised, is eliminated in the model activated sludge unit by > 90%. The elimination is calculated to be 97.75% taking into account the inflow into the model activated sludge unit of the test substance with a concentration of 10 mg/l, the dilution of the effluent of the elimination unit in ratio 1:1.25 and the separately determined EC0=0.1 mg/l to Daphnia magna. Thus a NOEC of >100 μg/L is calculated.

The test material contains isopropanol. The toxicity of isopropanol against Daphnia magna is more than 10 000 times lower than the toxicity of the test material. Water solubility of the test item may be slightly enhanced leading to a better bioavailability and slightly enhanced toxicity. But these effects are regarded to have a minor influence on the test result.Therefore it can be concluded in a first approach, neglecting additive effects, that the observed effect values can be attributed to the active ingredient itself.

This study is classified as acceptable and satisfies basic scientifically requirements for a chronic toxicity study with freshwater invertebrates, although not all data were reported.

Justification for read-across:

The structural similarities between the source and the target substances presented above are the basis for the read-across hypothesis. Adequate, reliable and available scientific information indicates that the source and target substances have similar physicochemical properties, ecotoxicological and toxicity profiles and thus support the read-across hypothesis.

Both substances are UVCB substances, produced by a similar process resulting in main constituents of the same structure, varying in the degree of saturation and chain length (C16 and C18 and mainly C18, for source and target substance, respectively). Given the underlying identical generic structure (outlined in chapter 1 and 2), similar absorption following oral or dermal uptake and the same metabolic patterns are expected for source and target substance. The findings from toxicokinetic data confirm that the discussed IQAC source and target substances are only poorly absorbed after oral application and rapidly excreted. There was no tendency for accumulation of the substance in the body of the test animals.

In conclusion the results obtained from source substance (partially unsaturated IQAC, DMS quaternised) are considered a reliable source to cover endpoints of the target substance (oleic-acid based IQAC, DMS quaternised). Beyond, the dose descriptors obtained from these studies performed on the source substance are considered as an appropriate starting point for deriving the respective PNECs.

A more detailed justification for read-across is outlined in a separate document:

“Justification for read-across - toxicological information”, is attached to the endpoint summary acute toxicity and provided in chapter 13 of Technical dossier.