1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-(C8-18(even numbered) and C18 unsaturated acyl) derivs., hydroxides, inner salts

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11.09.2007-15.09.2007

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

adapted by OSPARCOM (2005-11) for marine testing of offshore chemicals

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Coco AAPB was observed to be a soluble material and an appropriate quantity was added to 1000 mL of treated seawater, stirred on a bench-top stirrer for 1 h, 10 min and for 1 h, respectively. Homogenisation of the material was performed by gentle inversion and moderate shaking of the sample container prior to weighing into the mixing vessel.
- 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):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM
- Common name: sheepshead minnow (Cyprinodon variegatus)
- Source: Opus Plus Ltd. own breeding
- Age at study initiation (mean and range, SD): 175 d
- Length at study initiation (length definition, mean, range and SD): 1.5-2.5 cm
- Weight at study initiation (mean and range, SD): 0.193 g (mean)
- Method of breeding: Sheepshead minnow were held under flow-through conditions in 50 litre capacity glass tanks, containing 1 µm filtered, ultra-violet treated seawater and supplied with gentle aeration. The fish were fed twice daily with flakes ad libitum.
- Feeding during test: no

ACCLIMATION: not reported

Test type:

semi-static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

19.2-21.2°C

pH:

7.97-8.29

Dissolved oxygen:

98-100%

Salinity:

36°/oo

Nominal and measured concentrations:

3 mg product/L nominal

Details on test conditions:

TEST SYSTEM
- Test vessel: 15 L glass tank
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass/15 L/-/3 L
- Aeration: yes
- Type of flow-through (e.g. peristaltic or proportional diluter): test conducted under semi-static conditions
- Renewal rate of test solution (frequency/flow rate): the test media were replaced after 48 h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.642 g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: natural seawater was used in the test: for this natural local seawater was pumped to holding tanks from Scapa Flow, Orkney. The seawater was sand filtered to remove particulate, filtered to 1 µm, heated to 16°C, UV treated, and degassed. The water used in the facility had a salinity of 35-36°/oo.
- Culture medium different from test medium: no
- Intervals of water quality measurement: dissolved oxygen, pH, temperature and salinity were measured prior and following each renewal of media

OTHER TEST CONDITIONS
- Adjustment of pH: not reported in respect to the test substance
- Photoperiod: a cylcle of low level light graduating to a higher level was used
- Light intensity: not reported

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: only one concentration tested
- Justification for using less concentrations than requested by guideline: limit test
- Range finding study
- Test concentrations: 5 concentrations tested (not further specified)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

1.1 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

not specified

Conclusions:

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 for 96 h, no effects were observed at 3 mg product/L nominal (corresponding to 1.1 mg a.i./L nominal).

Executive summary:

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 for 96 h. No effects were observed at 1.1 mg a.i./L nominal (=3 mg product/L).

This study is regarded as reliable with restrictions.

Results Synopsis
Test organism/length/weight: Cyprinodon variegatus/18.3 mm (average)/0.193 g (average)
Test Type: semi-static
96 h NOEC: 1.1 mg a.i./L (nominal)

Endpoint(s) Effected: mortality

Reference 2

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

Description of key information

Reliable results on the acute toxicity of 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

Dose descriptor:

EC50

Effect concentration:

1.11 mg/L

Marine water fish

Marine water fish

Dose descriptor:

EC50

Effect concentration:

1.1 mg/L

Additional information

Short-term toxicity studies in fish were conducted with the target substance C8-18 and C18 unsatd. AAPB (Coco AAPB).

A prolonged fish toxicity test and a further short-term toxicity study in freshwater fish are available for the closely related source substance C8-18 AAPB. A justification for read-across is given below.

 

Freshwater fish

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.

 

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

 

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
	C12 AAPB	C8-18 AAPB	C8-18 and C18 unsatd. AAPB
Short-term toxicity to fish	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	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
		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/Ltest mat. (nominal) based on: mortality 96 h LC0 = 10.7 mg/Ltest 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/Lact. ingr. (nominal) based on: mortality 96 h LC0 = 1.7 mg/Lact. 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 substanceC8-18 AAPB are also valid for the target substance C8-18 and C18 unsatd. 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 be 1.11 mg a.i./L nominal, and for marine fish (Cyprinodon variegatus) 1.1 mg a.i./L nominal.