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Short-term toxicity to fish

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Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
23.02.1995-23.03.1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 215 Draft 'Juvenile growth test: 28 d' (1992)
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
yes
Details on sampling:
Stability of the test chemical solutions: concentration of stock solution was measured weekly
Vehicle:
no
Details on test solutions:
STOCK AND TEST SOLUTION AND THEIR PREPARATION
A stock solution (16 g/L nominal) was prepared for each test concentration every 2 to 4 days under sterile conditions at a pH of 5 and a temperature of 4°C.
Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Source: Forellenhof Fredersloh, D-37186 Moringen (Germany)
- Age at study initiation (mean and range, SD): young fish (not further specified)
- Length at study initiation (length definition, mean, range and SD): mean in all test vessels: 5.0-5.2 cm (range: 4.5-5.7 cm); SD: 0.26-0.33
- Weight at study initiation (mean and range, SD): mean in all test vessels: 1.4-1.5 g ww (range: 1.2-2.1 g); SD: 0.14-0.29
- Feeding during test
- Food type: commercial trout diet
- Amount: daily 4% dry weight related to the mean initial fish wet weight; the food quantity was weekly increased during the test by 25% related to the initial food ration respectively to the food ration of the week before
- Frequency: daily

ACCLIMATION
- Acclimation period: >4 weeks
- Acclimation conditions (same as test or not): same
- Type and amount of food: commercial trout diet; daily 4% dry weight related to the mean initial fish wet weight
- Feeding frequency: daily
- Health during acclimation (any mortality observed): yes
Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d
Hardness:
2.00-2.25 mmol/L=200-225 mg/L as CaCO3
Test temperature:
13.0-15.1°C
pH:
7.3-8.0
Dissolved oxygen:
8.0-9.8 mg/L
Nominal and measured concentrations:
Concentrations:
0.05, 0.16, 0.50, 1.6 and 5.0 mg/L product (nominal)
0.016, 0.05, 0.16, 0.5 and 1.6 mg/L active substance (nominal)
Details on test conditions:
TEST SYSTEM
- Test vessel: glass aquarium
- Type (delete if not applicable): not reported
- Material, size, headspace, fill volume: glass, 0.54x0.29x0.395 m
- Aeration: not reported
- Type of flow-through (e.g. peristaltic or proportional diluter): HAMILTON Digital dispenser
- Renewal rate of test solution (frequency/flow rate): renewal rate: eightfold theoretical volume exchange/d; flow rate: 16 L/h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): no vehicle used
- Biomass loading rate: ca. 0.3 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking water (total hardness was reduced and the water filtered through an activated carbon filter)
- Alkalinity: 4.4-4.7 mmol/L
- Culture medium different from test medium: no
- Intervals of water quality measurement:: total hardness and alkalinity was measured 5 times weekly; temperature, pH, and dissolved oxygen were measured daily

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16 h light/8 h dark
- Light intensity: 10-25 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality (daily) and growth (at test start and test end)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 3
- Range finding study: performed
- Results used to determine the conditions for the definitive study: yes
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
ca. 0.16 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Duration:
28 d
Dose descriptor:
LOEC
Effect conc.:
ca. 0.5 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Details on results:
Nominal/measured concentrations: nominal
- Mortality:
< 1.6 mg/L: no deaths
>= 1.6 mg/L: 10/10 fish died (within 4 days)

- Growth (wet weight, length):
<= 0.5 mg/L: no significant differences

- Controls:
length: increase with factor 1.4;
wet weight: increase with factor 2.9

- Effect concentrations:
all values relate to nominal concentrations
28 d NOEC= 0.5 mg product/L
28 d LOEC= 1.6 mg product/L
28 d LLC= 1.6 mg product/L (LLC: lowest lethal concentration)

Assuming a content of 32.1% active substance:
28 d NOEC= 0.16 mg active substance/L
28 d LOEC= 0.5 mg active substance/L
28 d LLC= 0.5 mg active substance/L
Reported statistics and error estimates:
ANOVA, Williams
Validity criteria fulfilled:
yes
Conclusions:
In a 28-d toxicity study conducted according to OECD Guideline 204 under flow-through conditions and using rainbow trouts (Oncorhynchus mykiss) as test organism, the 28-d NOEC and LOEC were 0.16 and 0.5 mg a.i./L nominal based on mortality and sublethal effects.
Executive summary:

In a 28-d toxicity study conducted according to OECD Guideline 204, rainbow trouts (Oncorhynchus mykiss) were exposed to C8 -18 AAPB at nominal concentrations of 0.016, 0.05, 0.16, 0.5 and 1.6 mg a.i./L (= 0.05, 0.16, 0.50, 1.6 and 5.0 mg/L product (nominal)) under flow through conditions.  The 28-d NOEC and LOEC were 0.16 and 0.5 mg a.i./L nominal based on mortality and sublethal effects. The validity criteria of the OECD Guideline 204 were fulfilled.

This study is classified as reliable without restriction.

Results Synopsis

Test organism/age/size: Oncorhynchus mykiss/-/4.5 - 5.7 cm; 1.2 - 2.1 g wwt at start of test

Test Type: Flow-through

28 d NOEC:  0.16 mg a.i./L (nominal)

28 d LOEC: 0.5 mg a.i./L (nominal)

Endpoint(s) Effected: mortality and sublethal effects

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
30.03.1992-03.04.1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
GLP compliance:
not specified
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: 6.67 g of the test material were dissolved in 2 L of distilled water to give a 1 g a.i./L stock. A range of test solution concentrations was prepared by diluting the 1 g/L stock with carbon-filtered tap water. Test solutions were prepared in 20 litre volumes and renewed every 24 h.
- Controls: performed
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): no vehicle used
- Evidence of undissolved material (e.g. precipitate, surface film, etc): not reported
Test organisms (species):
Pimephales promelas
Details on test organisms:
TEST ORGANISM
- Common name: fathead minnow
- Source: Unilever own breeding
- Age at study initiation (mean and range, SD): not reported
- Length at study initiation (length definition, mean, range and SD): not reported (length of fish in the control at the end of the test: 51 mm)
- Weight at study initiation (mean and range, SD): not reported (wet weight of fish in the control at the end of the test: 1.9 g)
- Method of breeding: The fish were reared in carbon-filtered tap water and fed on proprietary trout food (no futher details reported).
- Feeding during test: no

ACCLIMATION
- Acclimation period: 24 h
- Acclimation conditions (same as test or not): not reported
- Type and amount of food: trout food (no futher details reported)
- Feeding frequency: not reported
- Health during acclimation (any mortality observed): no significant mortalities of the test fish in the week prior to the test
Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Hardness:
total hardness as mg/L CaCO3: 78-105
Test temperature:
22.5-23.0°C
pH:
7.2-7.6
Dissolved oxygen:
7.1-8.7 mg/L
Nominal and measured concentrations:
0 (control), 1.0, 1.8, 3.2, 5.6, and 10 mg product/L nominal
0 (control), 0.3, 0.54, 0.96, 1.68, and 3.0 mg a.i./L nominal
Details on test conditions:
TEST SYSTEM
- Test vessel: 30 L test vessels
- Type (delete if not applicable): not reported
- Material, size, headspace, fill volume: -/30 L/-/20 L
- Aeration: not reported
- Type of flow-through (e.g. peristaltic or proportional diluter): test conducted under semi-static conditions
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: ca. 0.6 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water
- Culture medium different from test medium: not reported
- Intervals of water quality measurement: temperature, DO, pH and total hardness were measured daily

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light/8 h dark
- Light intensity: not measured (normal room lighting was used)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality, behaviour

TEST CONCENTRATIONS
- Spacing factor for test concentrations: ca. 1.73
- Range finding study: not reported
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
0.54 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
1.11 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% c.i.: 0.54-1.68 mg a.i./L
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
1.68 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Reported statistics and error estimates:
The effect concentrations were calculated by binominal analysis.
Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of Coco AAPB to fish (Pimephales promelas) was investigated in a study conducted according to OECD Guideline 203 and EU Method C.1. A 96 h LC50=1.11 mg a.i./L nominal (=3.7 mg product/L) was determined.
Executive summary:

The acute toxicity of Coco AAPB to fish (Pimephales promelas) was investigated in a study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test) and EU Method C.1 (Acute Toxicity for Fish). The fish were exposed to nominal concentrations of 0 (control), 1.0, 1.8, 3.2, 5.6, and 10 mg product/L (0 (control), 0.3, 0.54, 0.96, 1.68, and 3.0 mg a.i./L) for 96 h under semi-static conditions. Based on the results a 96 h LC50=1.11 mg a.i./L nominal (=3.7 mg product/L) was calculated via binominal analysis. Observations of the behaviour of the fish during the test indicated that behaviour was affected but not at concentrations significantly lower than the 96 h LC50.

This study is regarded as reliable without restriction.

Results Synopsis
Test organism/length/weight: Pimephales promelas/ca. 50 mm/1.9 g

Test Type: semi-static

96 h LC50: 1.11 mg a.i./L (nominal)

Endpoint(s) Effected: mortality

Endpoint:
short-term toxicity to fish
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Justification for type of information:
REPORTING FORMAT FOR THE ANALOGUE APPROACH
see "General Justification for Read-Across" attached to IUCLID section 13

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
Mutual read across from the AAPBs to one another is justified:

a) Based on the information given in section 1, it can be concluded that all AAPBs mentioned above are similar in structure, since they are manufactured from similar resp. identical precursors under similar conditions and all contain the same functional groups. Thus a common mode of action can be assumed.
b) The content of minor constituents in all products are comparable and differ to an irrelevant amount.
c) The only deviation within this group of substances is a minor variety in their fatty acid moiety, which is not expected to have a relevant impact on intrinsic toxic or ecotoxic activity and environmental fate. Potential minor impact on specific endpoints will be discussed in the specific endpoint sections.

The read-across hypothesis is based on structural similarity of target and source substances. Based on the available experimental data, including key physico-chemical properties and data from toxicokinetic, acute toxicity, irritation, sensitisation, genotoxicity and repeated dose toxicity studies, the read-across strategy is supported by a quite similar toxicological profile of all five substances.
The respective data are summarised in the data matrix; robust study summaries are included in the Technical Dossier in the respective sections.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
see "General Justification for Read-Across" attached to IUCLID section 13

3. ANALOGUE APPROACH JUSTIFICATION
see "General Justification for Read-Across" attached to IUCLID section 13

4. DATA MATRIX
see "General Justification for Read-Across" attached to IUCLID section 13
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across source
Reason / purpose for cross-reference:
read-across: supporting information
Key result
Duration:
96 h
Dose descriptor:
LC50
Remarks:
(freshwater fish: Pimephales promelas)
Effect conc.:
1.11 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Remarks on result:
other: 95% c.i.: 0.54-1.68 mg a.i./L
Key result
Duration:
96 h
Dose descriptor:
NOEC
Remarks:
(marine fish: Cyprinodon variegatus)
Effect conc.:
1.1 mg/L
Nominal / measured:
nominal
Conc. based on:
act. ingr.
Basis for effect:
mortality (fish)
Conclusions:
For freshwater fish (Pimephales promelas) the lowest 96 h LC50 was determined to be 1.11 mg a.i./L nominal, and for marine fish (Cyprinodon variegatus) 1.1 mg a.i./L nominal.
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
February 17, 2020 - May 18, 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Version / remarks:
adopted June 2019
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
- Chemical name (IUPAC): 1-Propanaminium, N-(carboxymethyl)-N,N-dimethyl-3-[(1-oxododecyl)amino]-, inner salt
Analytical monitoring:
yes
Details on sampling:
Samples from test solutions (fresh and aged) were taken for analysis of actual test item concentrations in comparison to the nominal concentrations.
Each sample contained 20 mL of test solution. The samples were stored in amber glass bottles in a freezer in the dark at a temperature of ≤ -18°C and shipped on dry ice.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
At start of the test (day 0) and at day 2 stock solutions at a nominal concentration of 80.0 mg test item/L were prepared by adding 402.0 mg test item to 5000 mL (day 0) and 241.91 mg test item to 3000 mL (day 2) test medium. These stock solutions were stirred for 30 min at 400 rpm on a magnetic stirrer at ambient temperature. These stock solutions were used to prepare the desired test item concentrations and were used immediately after preparation.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Source: stock culture (ECT)
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): 1.20–1.95 cm, mean: 1.62 cm (assessed from the control and all treatments at end of the exposure period or at time of removal of dead fish)
- Weight at study initiation (mean and range, SD): range: 0.0270–0.0491 g/fish, mean: 0.0371 g/fish (assessed from a representative sub-sample consisting of 10 fish, three days before start of exposure)
- Maintenance of the brood fish:
− Amount of water per stock vessel: approximately 25 L
− Depth of water in the stock vessel: approximately 21 cm
− Estimated number of fish kept as a stock per holding vessel (batch size): approx. 325


ACCLIMATION
- Acclimation conditions (same as test or not): same
- Type and amount of food during acclimation: TetraMin®, Artemia nauplii and Paramecium caudatum
- Feeding frequency during acclimation: daily ration fed in 3 portions on workdays and one portion on weekend, ad libitum, no feeding on the day (>24 hours) before start of exposure
- Health during acclimation (any mortality observed): Mortalities during the period of 7 d before start of the test (<5%): 0%




Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Test temperature:
22.8 - 23.6°C
pH:
7.4 - 7.7
Dissolved oxygen:
8.0 - 8.9 mg/L (95 - 105% of air saturation)
Nominal and measured concentrations:
nominal 7.62, 13.7, 24.7, 44.4 and 80.0 mg test item/L, equivalent to 2.80, 5.03, 9.08, 16.3, 29.4 mg solids/L (corrected for water content of 63.25%)
Details on test conditions:
TEST SYSTEM
- Test vessel: 3.8 L glass vessels covered by transparent lids, 2 L fill volume
- Aeration: permanent
- Renewal rate of test solution (frequency/flow rate): renewal after 48 h
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.130 g/L (assessed from the control fish at end of the exposure period)

TEST MEDIUM / WATER PARAMETERS
Reconstituted water according to OECD guideline No. 203


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 14 h light / 10 h dark
- Light intensity: 540–1000 lx; measured: 540–572 lx, (n = 6)

EFFECT PARAMETERS MEASURED :
2, 5, 24, 29, 48, 53, 72 and 96 hours after start of exposure for lethality + sublethal effect

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
49.4 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
18.2 other: mg solids/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Other biological observations: A clear concentration-response relationship was observed. Sub-lethal effects (behavioural impairments) were observed down to the intermediate treatment level (C3, 24.7 mg test item/L). No sub-lethal effects were observed in the control. Concluding from the empirical evaluation of these observations, a NOECbehavioural impairments of 24.7 mg test item/L based on nominal concentrations can be derived from this test
- Mortality of control: 0
Results with reference substance (positive control):
n.a.
Reported statistics and error estimates:
The data were evaluated by the qualitative trend analysis by contrasts (monotonicity of concentration/response). Mortality of the fish was evaluated. Probit analysis using linear maximum likelihood regression was used to estimate the LCx values. The No-Observed Effect Concentration (NOEC) and the Lowest Obsverved Effect Concentration (LOEC) were evaluated by the Stepdown Cochran-Armitage Test Procedure.
The statistical software package ToxRat Professional 3.3.0 (ToxRat Solutions GmbH, Naheweg 15, D-52477 Alsdorf) was used for these calculations. The statistical analysis was performed using nominal concentrations.
Sublethal observations / clinical signs:

All validity criteria were fulfilled as required by the study plan:

Mortality in the control: (required: </= 10%, or one out of seven fish) 0%

Dissolved oxygen concentration in control and test vessels: 8.0–8.9 mg/L (required: >/= 60% of air saturation value (ASV)) (ASV: >/= 100%)

Analytically Determined Test Item Concentrations

The concentration measured in the freshly prepared test solutions at the start (0 hours) and at 48 hours (day 2) of the exposure ranged from 96–102% of the nominal concentrations and remained stable during each renewal period of 48 hours within a range of 88–99% of the nominal concentrations. One exception was observed in an aged test solution at the lowest concentration level after 96 hours, for which a decrease was observed down to a recovery of 44% of nominal concentration.

Note: Since the measured test item concentrations at biological relevant treatment levels (i.e. C3, C4 and C5; 24.7, 44.4 and 80.0 mg test item/L) showed stable concentration within ±20% of nominal concentration at all test solution renewal intervals (0–48 and 48–96 hours) the low recovery at the treatment level (C1) was considered to be an outlier and was not taken into account while evaluation the biological results.

Therefore, the measured concentrations at the biological relevant concentration levels were within ±20% of the nominal concentrations, and the biological data were evaluated based on nominal concentration expressed as mg test item/L, and in addition expressed in mg solids/L.

Cumulative mortality of the fish

 

Nominal concentrations

[mg test item/L]

 

Control

7.62

13.7

24.7

44.4

80.0

 

Time [hours]

Mortality

0

0

0

0

0

0

0

2

0

0

0

0

0

0

5

0

0

0

0

0

7

24

0

0

0

0

0

7

29

0

0

0

0

0

7

48

0

0

0

0

0

7

53

0

0

0

0

0

7

72

0

0

0

0

1

7

96

0

0

0

0

1

7

Observations on behaviour and appearance of the fish.

Nominal concentrations

[mg test item/L]

 

Control

7.62

13.7

24.7

44.4

80.0

 

Time [hours]

Observations

0

-

-

-

-

-

-

2

-

-

-

-

-

-

5

-

-

-

-

E5, L7

E7, L7

24

-

-

-

E1, L1

A3, E5, L7

n.a.

29

-

-

-

-

A7, E7, L7

n.a.

48

-

-

-

-

 

n.a.

53

-

-

-

-

E6

n.a.

72

-

-

-

-

E6

n.a.

96

-

-

-

-

-

n.a.

 

A: fish are lying on the bottom of the test vessel

E: fish show reduced swimming activities compared to the control fish

L: fish are only swimming near by the bottom of the test vessel

n.a.: not applicable, all fish were dead at this observation time.

Validity criteria fulfilled:
yes
Conclusions:
The acute toxicity of C12 AAPB to zebrafish (Danio rerio) was investigated in a study conducted according to OECD Guideline 203. A 96 h LC50 of 49.4 mg/Las test item, corresponding to 18.2 mg solids/L was determined.
Executive summary:

A study on the acute toxicity of C12 AAPB to zebrafish (Danio rerio) was conducted according to OECD Guideline 203 (adopted June 2019).

The fish (7/concentration) were exposed to nominal concentrations of 0 (control),7.62, 13.7, 24.7, 44.4 and 80.0 mg test item/L, which correspond to the following concentrations of 2.80, 5.03, 9.08, 16.3, 29.4 mg solids/ L, for 96 h under semi-static conditions.

The concentration measured in the freshly prepared test solutions ranged from 96102% of the nominal concentrations and remained stable during each renewal period of 48 hours within a range of 8899% of the nominal concentrations.

Since the measured concentrations were in most cases within ±20% of the nominal concentrations, the biological data were evaluated based on nominal concentration expressed as mg test item/L and mg solids/L.

A clear concentration-response relationship was observed. The following LC50 and NOEC/LOEC values were determined based on statistical evaluation of biological results and nominal concentrations of the test item:

96 h LC50 =49.4 mg/Las test item, corresponding to 18.2 mg solids/L

NOEC = 44.4 mg/L as test item,corresponding to 16.3 mg solids /L

LOEC =80 mg/L as test item,corresponding to 29.4 mg solids /L

Description of key information

Reliable results on the acute toxicity of C12 AAPB, C8-18 AAPB and Coco AAPB and the prolonged toxicity of C8-18 AAPB to fresh water fish obtained from guideline studies are available. For marine fish a limit test conducted according to guideline is available.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish
Effect concentration:
1.11 mg/L

Marine water fish

Marine water fish
Effect concentration:
1.1 mg/L

Additional information

Short-term toxicity studies in freshwater and saltwater fish are available for C12 AAPB as well as for the closely related source substances C8-18 AAPB and C8-18 and C18 unsatd. AAPB (Coco AAPB).A justification for read-across is given below.

 

Freshwater fish

Tests with C12 AAPB:

A study on the acute toxicity of C12 AAPB to zebrafish (Danio rerio) was conducted according to OECD Guideline 203 (adopted June 2019).

The fish (7/concentration) were exposed to nominal concentrations of 0 (control),7.62, 13.7, 24.7, 44.4 and 80.0 mg test item/L, which correspond to the following concentrations of 2.80, 5.03, 9.08, 16.3, 29.4 mg solids/ L, for 96 h under semi-static conditions.

The concentration measured in the freshly prepared test solutions ranged from 96–102% of the nominal concentrations and remained stable during each renewal period of 48 hours within a range of 88–99% of the nominal concentrations.

Since the measured concentrations were in most cases within ±20% of the nominal concentrations, the biological data were evaluated based on nominal concentration expressed as mg test item/L and mg solids/L.

A clear concentration-response relationship was observed. The following LC50 and NOEC/LOEC values were determined based on statistical evaluation of biological results and nominal concentrations of the test item:

96 h LC50 =49.4 mg/Las test item, corresponding to 18.2 mg solids/L

NOEC = 44.4 mg/L as test item, corresponding to 16.3 mg solids /L

LOEC =80 mg/L as test item, corresponding to 29.4 mg solids /L

 

Tests with Coco AAPB:

In a semi-static test conducted according to OECD Guideline 203 and EU Method C.1 usingPimephales promelas as test organism a 96 h LC50=1.11 mg a. i./L (nominal; no analytical monitoring of test substance concentration; pH not adjusted; pH range: 7.2 -7.6) was determined. In two semistatic tests conducted according to OECD Guideline 203 and/or EC Method C.1 with Danio rerio (pH adjusted) and Cyprinus carpio (pH not adjusted; pH range: 7.8 -8.2) 96 h LC50 values of ca. 2 mg a. i./L (nominal) and 1.9 mg a.i./L (nominal), respectively, were derived. Analytical monitoring of test substance concentrations was not conducted in the test with Danio rerio. In the semi-static test with Cyprinus carpio the concentration of Coco AAPB was stable during the incubation time which has been shown by analytical monitoring via TOC (non-specific method).

 

Tests with C8-18 AAPB:

The acute toxicity of C8-18 AAPB towards fish (Danio rerio) was investigated in a study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test) and ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae) ] - Part 1: Static Method). The 96 h LC50 was found to be 15 mg/L nominal (no information whether this value refers to test material or active matter).

Furthermore the results of a prolonged fish toxicity test are available. In this 28-d toxicity study conducted according to OECD Guideline 204, rainbow trouts (Oncorhynchus mykiss) were exposed to C8-18 AAPB under flow-through conditions. The 28-d NOEC and LOEC values were 0.16 and 0.5 mg a. i./L nominal based on mortality and sub-lethal effects.

 

Saltwater fish

The acute toxicity of Coco AAPB to the marine fish Cyprinodon variegatus was investigated in a study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test) adapted by OSPARCOM (2005 -11) for marine testing of offshore chemicals. In this limit test conducted under semi-static conditions, the fish were exposed to a single concentration of 3 mg product/L nominal (corresponding to 1.1 mg a. i./L) for 96 h. No effects were observed at 1.1 mg a. i./L nominal.

 

Conclusion

For freshwater fish (Pimephales promelas) the lowest 96 h LC50 was determined to be 1.11 mg a.i./L nominal, and for marine fish (Cyprinodon variegatus) 1.1 mg a.i./L nominal.

The obtained results were considered to be valid for the AAPBs.

 

 

Justification for read-across

For details on substance identity and detailed (eco)toxicological profiles, please refer also to the general justification for read-across given at the beginning of the CSR and attached as pdf document to IUCLID section 6.1 (Aquatic toxicity).

 

This read-across approach is justified based on structural similarities. All AAPBs contain the same functional groups. Thus a common mode of action can be assumed.

The only deviation within this group of substances is a minor variety in their fatty acid moiety (chain length and degree of unsaturation), which is not expected to have a relevant impact on intrinsic ecotoxicological properties.

 

a. Structural similarity and functional groups

Alkylamidopropyl betaines (AAPBs) are – with the exception of C12 AAPB - UVCB substances (Substances of Unknown or Variable composition, Complex reaction products or Biological materials), which are defined as reaction products of natural fatty acids or oils with dimethylaminopropylamine and further reaction with sodium monochloroacetate. AAPBs are amphoteric surfactants, which are characterized by both acidic and alkaline properties.

 

Their general structure is:

 

R-C(O)-NH-(CH2)3-(N(CH3)2)+-CH2-C(O)O-

R = fatty acid moiety

 

The fatty acids have a mixed, slightly varying composition with an even numbered chain length from C8 to C18. Unsaturated C18 may be included. Consequently, the AAPBs differ by their carbon chain length distribution and the degree of unsaturation in the fatty acid moiety. However, Lauramidopropyl betaine (C12 fatty acid derivate) is the major ingredient of all AAPBs covered by this justification as listed in table 1 “Substance identities” of the general justification for read-across.

 

The substances under evaluation share structural similarities with common functional groups (quaternary amines, amide bonds and carboxymethyl groups), and fatty acid chains with differences in chain length and degree of saturation.

 

b. Differences

Differences in acute ecotoxicity of the AAPBs could potentially arise from the following facts:

-Different amounts of different carbon chain lengths (carbon chain length distribution):

Higher amounts of higher chain lengths and corresponding lower amounts of lower chain length could result in a rising average lipophilicity. However, the main component for all AAPBs is C12 AAPB. Relevant effects on ecotoxicity are not to be expected.

- Different amounts of unsaturated fatty ester moieties:

Effects may be expected for e.g. physical state, but are not considered to be of relevance for ecotoxicity.

 

Comparison of short-term fish toxicity data

 

Endpoints

Source substances

Target substance

 

C8-18 AAPB

C8-18 and C18 unsatd. AAPB

C12 AAPB

Short-term toxicity to fish

key(prolonged fish test: 28 d).Short-term toxicity to fish.97862-59-4_9.1.3_IUA_1995_OECD_204

key study


OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study)

 

Oncorhynchus mykiss, flow-through, freshwater

 

28 d NOEC = ca. 0.16 mg/L act. ingr. (nominal) based on: mortality

 

28 d LOEC = ca. 0.5 mg/L act. ingr. (nominal) based on: mortality

 

No deaths < 1.6 mg/L; 10/10 fish died within 4 d at >/= 1.6 mg/L

 

Reliability: 1 (reliable without restriction), GLP

 

key.Short-term toxicity to fish.61789-40-0_9.1.3_Acute_fish_Unilever_AT-R728-01


key study

OECD Guideline 203 (Fish, Acute Toxicity Test)

Pimephales promelas, semi-static, freshwater

96 h NOEC = 0.54 mg/L,act. ingr. (nominal) 


96 h LC100 = 1.68 mg/L,
act. ingr. (nominal) 


96 h LC50 = 1.11 mg/L,
act. ingr. (nominal) based on: mortality (95% c.i.: 0.54-1.68 mg a.i./L)

 

Reliability: 1 (reliable without restriction), no GLP

 

 

key_Short-term toxicity to fish: 4292-10-8_9.1.3._Solvay_2020_OECD203

 

key study

OECD Guideline 203 (Fish, Acute Toxicity Test)

Danio rerio, semi-static, freshwater

 

96 h LC50 = 49.4 mg/Las test item, corresponding to 18.2 mg solids/L

 

Reliability: 1 (reliable without restriction), GLP

 

sup.Short-term toxicity to fish.97862-59-4_9.1.3_EOC_Lovap_1995_OECD 203

 

supporting study

 

OECD Guideline 203 (Fish, Acute Toxicity Test)

 

Brachydanio rerio (new name: Danio rerio), static, freshwater

96 h LC50 = 15 mg/Ltest mat. (nominal) based on: mortality


96 h LC100 = 21.5 mg/L
test mat. (nominal) based on: mortality


96 h LC0 = 10.7 mg/L
test mat. (nominal) based on: mortality

 

Reliability: 2 (reliable with restrictions), no GLP

 

key (marine).Short-term toxicity to fish.61789-40-0_9.1.3_Marine_Cyprinodon variegatus_Rhodia_Mirataine-BET-C-37_OECD203_2008


key study

 

similar to OECD Guideline 203 (Fish, Acute Toxicity Test) (adapted by OSPARCOM (2005-11) for marine testing of offshore chemicals)


Cyprinodon variegatus, semi-static, saltwater

 

96 h NOEC = 1.1 mg/Lact. ingr. (nominal) based on: mortality

 

Reliability: 2 (reliable with restrictions), GLP

 

 

sup.Short-term toxicity to fish: 61789-40-0_9.1.3_Henkel_2001_OECD 203


supporting study

 

OECD Guideline 203 (Fish, Acute Toxicity Test)


Brachydanio rerio (new name: Danio rerio), semi-static, freshwater

 

96 h LC100 = ca. 2.4 mg/Lact. ingr. (nominal) based on: mortality


96 h LC50 = ca. 2 mg/L
act. ingr. (nominal) based on: mortality


96 h LC0 = 1.7 mg/L
act. ingr. (nominal) based on: mortality

 

Reliability: 2 (reliable with restrictions), no GLP

 

sup.Short-term toxicity to fish: 61789-40-0_9.1.3_Hüls_1996_EEC 92-69 C1

 

supporting study


EU Method C.1 (Acute Toxicity for Fish)

Cyprinus carpio, semi-static, freshwater

96 h LC50 = 1.9 mg/Lact. ingr. (nominal) based on: mortality

 

96 h LC0 = 1.4 mg/Lact. ingr. (nominal) based on: mortality 

 

96 h LC100 = 2.5 mg/Lact. ingr. (nominal) based on: mortality

 

Reliability: 2 (reliable with restrictions), GLP

 

RL4_Short-term toxicity to fish: 61789-40-0_9.1.3_THG_1980c_DIN 38 412_48h

 

Supporting study

 

DIN 38412

Leuciscus idus

 

48 h LC0 = 2.5 mg/L based on: mortality

 

48 h LC100 = 5 mg/L based on: mortality

 

Reliability: 4 (not assignable), no GLP

 

LC50 values were in the range from 1.1 to 15 mg/L for different species.

For freshwater fish (Pimephales promelas) the lowest 96 h LC50 was determined to be 1.11 mg a.i./L nominal. For marine fish (Cyprinodon variegatus) the lowest 96 h LC50 was 1.1 mg a.i./L nominal.

 

Quality of the experimental data of the analogues:

The available data are adequate and sufficiently reliable to justify the read-across approach.

Two key studies with freshwaster fish have been identified, which were conducted according toOECD Guideline 204 (C8-18 AAPB, RL1, GLP) andOECD Guideline 203 (C8-18 and C18 unsatd. AAPB, RL1, no data on GLP), respectively.

A further key study with marine fish was conducted similar to OECD Guideline 203 (C8-18 and C18 unsatd. AAPB, RL2, GLP)

One supporting study conducted withC8-18 AAPB (RL2) and two supporting studies conducted withC8-18 and C18 unsatd. AAPB(RL 2) according toEU Method C.1 or OECD Guideline 203 are reported.

The test materials used in the respective studies represent the source substance as described in the hypothesis in terms of substance identity and minor constituents.

Overall, the study results are adequate for the purpose of classification and labelling and risk assessment.

 

Conclusion

Based on structural similarities of the target and source substancesas presented above and in more detail in the general justification for read across, it can be concluded that the available data from the source substancesC8-18 AAPB and C8-18 and C18 unsatd. AAPB are also valid for the target substance C12 AAPB.

 

The obtained LC50 values were in the same range for freshwater and marine fish:For freshwater fish (Pimephales promelas) the lowest 96 h LC50 was determined to 1.11 mg a.i./L nominal, and for marine fish (Cyprinodon variegatus) to 1.1 mg a.i./L nominal.