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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16.10.1990-18.10.1990

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

MONITORING OF TEST SUBSTANCE CONCENTRATION:
Analytical monitoring at the beginning and the end of test.

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: waterflea (Daphnia magna)
- Source: RCC own breeding
- Age at study initiation (mean and range, SD): <24 h
- Method of breeding: Breeding was performed in about 10 L glass vessels in mixtures of pond and reconstituted water with slight aeration. They were fed with algae and kept under the same temperature and light conditions as used in the test itself.
- Feeding during test: no

ACCLIMATION
- Acclimation period: not necessary
- Acclimation conditions (same as test or not): yes

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.0 - 20.5 °C

pH:

8.3 - 8.4 (start), 8.2 - 8.4 (end)

Dissolved oxygen:

8.0 - 8.1 mg/L (start), 7.9 - 8.3 mg/L (end)

Nominal and measured concentrations:

Concentrations:
6.25; 12.5; 25; 50; 100 mg product/L (nominal)
1.9, 3.75, 7.5, 15, 30 mg active substance/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL beaker
- Material, size, headspace, fill volume: glass/50 mL/-/20 mL
- Aeration: not reported
- Type of flow-through (e.g. peristaltic or proportional diluter): test performed under static conditions
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted water prepared acc. to EEC Directive
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH and oxygen were measured at the beginning and at the end of the test

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h illumination
- Light intensity: 500-2000 Lux

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: performed (test conditions/concentrations not specified in this report)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

6.5 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95% C.I.: 4.8 - 8.4 mg a.i./L

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 30 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Details on results:

RESULTS: EXPOSED
- Nominal/measured concentrations: nominal
48h-EC0: 5.3 mg product/L
48h-EC50: 21.5 mg product/L
48h-EC100: 89.3 mg product/L

Assuming a content of 30% active substance:
48h-EC0: ca. 1.6 mg active substance/L
48h-EC50: ca. 6.5 mg active substance/L
48h-EC100: ca. 26.8 mg active substance/L

Reported statistics and error estimates:

The 48 h EC50 was estimated using the Logit-model and EC0, EC50, and EC100 values were determined by linear regression of values used for the Logit estimation.

The acute toxicity of C8 -18 AAPB to Daphnia magna was investigated in a study conducted according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) and EU Method C.2 (Acute Toxicity for Daphnia). The results were compiled in the following table. Table 1: Effect of TEGO Betain on immobilisation of Daphnia magna

Treatment (mg a.i./l) [nominal conc. ]		No. of organ-isms at test begin	Observation period
			24 h		48 h
			Cumul. no. of dead	% morta-lity	Cumul. no. of dead	% morta-lity
Control (dilution water only)	Replicate #1	10	0	0	0	0
	Replicate #2	10	0	0	0	0
1.9	Replicate #1	10	0	0	1	10
	Replicate #2	10	0	0	0	0
3.75	Replicate #1	10	0	0	3	30
	Replicate #2	10	0	0	1	10
7.5	Replicate #1	10	4	40	7	70
	Replicate #2	10	1	10	8	80
15	Replicate #1	10	1	10	8	80
	Replicate #2	10	3	30	7	70
30	Replicate #1	10	4	40	10	100
	Replicate #2	10	5	50	10	100

48h-EC0: 5.3 mg product/L

48h-EC50: 21.5 mg product/L

48h-EC100: 89.3 mg product/L

Assuming a content of 30% active substance:

48h-EC0: ca. 1.6 mg active substance/L

48h-EC50: ca. 6.5 mg active substance/L

48h-EC100: ca. 26.8 mg active substance/L

Validity criteria fulfilled:

yes

Conclusions:

The 48 h EC50 of C8-18 AAPB to Daphnia magna was determined to be 6.5 mg a.i./L nominal in a study conducted according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) and EU Method C.2 (Acute Toxicity for Daphnia).

Executive summary:

The 48 hr-acute toxicity of C8 -18 AAPB (30% active matter) to Daphnia magna was investigated in a study conducted according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) and EU Method C.2 (Acute Toxicity for Daphnia) under static conditions. Daphnids were exposed to control and test chemical at nominal concentrations of 6.25; 12.5; 25; 50; 100 mg product/L for 48 hr. The test material concentrations were analysed via UV absorption at 485 nm. The C8 -18 AAPB concentrations were >80% (exception: 72.2% at the start in the 6.25 mg product/L group). The 48 h EC50 was determined to be 6.5 mg a.i./L.

This study is classified as reliable without restriction.

Results Synopsis
Test Organism/Age (e.g. 1^st instar): Daphnia magna/<24h
Test Type: Static
48 h EC50:  6.5 mg a.i./L; 95% C.I.: 4.8 to 8.4 mg a.i./L nominal
Endpoint(s) Effected:  Immobilization

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

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

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EC50

Remarks:

freshwater invertebrates

Effect conc.:

ca. 1.9 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 95% CL: 1.37 - 2.69 mg a.i./L

Key result

Duration:

48 h

Dose descriptor:

LC50

Remarks:

marine invertebrates

Effect conc.:

7 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Remarks on result:

other: 95% c.i.: 5.4 - 8.3 mg a.i./L

Conclusions:

For freshwater invertebrates (Daphnia magna) the lowest 48 h EC50 was determined to be 1.9 mg a. i./L (nominal), and for the marine copepod Acartia tonsa  the 48 h EC50 was 7.0 mg a.i./L (nominal).

Description of key information

The acute toxicity of C8 -18 and C 18 unsatd. AAPB to Daphnia magna was investigated in four studies conducted according to OECD guideline 202. The study with the lowest 48 h EC50 was selected as key study. Additional data were available for C8-18 AAPB from an OECD 202 guideline study with Daphnia magna. The acute toxicity of Coco AAPB to the marine copepod Acartia tonsa was investigated in a study conducted according to ISO 14669 (1999) Water 
Quality - Determination of the acute lethal toxicity to marine capepods (Copepoda; Crustacea).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1.9 mg/L

Marine water invertebrates

Marine water invertebrates

Effect concentration:

7 mg/L

Additional information

Reliable results on the acute toxicity to fresh water invertebrates obtained in guideline studies are available for C8-18 AAPB and the closely related source substances C8-18 and C18 unsatd. AAPB and C12 AAPB, as well as data on the acute toxicity of C8-18 and C18 unsatd. AAPB to salt water invertebrates. A justification for read-across is given below.

 

Freshwater invertebrates

The acute toxicity of Coco AAPB to Daphnia magna was investigated in a study conducted according to OECD TG 202 / part I (1984) under static conditions. Daphnia magna was exposed to 6 nominal concentrations ranging from 0.5 to 16 mg product/L. A 48 h LC50 of ca. 1.9 mg a.i/L (nominal) was determined based on an active matter content of 30%.

 

The 48 hr-acute toxicity of C8-18 AAPB (30% active matter) to Daphnia magna was investigated in a study conducted according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) and EU Method C.2 (Acute Toxicity for Daphnia) under static conditions. Daphnids were exposed to control and test chemical at nominal concentrations of 6.25; 12.5; 25; 50; 100 mg product/L for 48 hr. The test material concentrations were analysed via UV absorption at 485 nm. The C8-18 AAPB concentrations were >80% (exception: 72.2% at the start in the 6.25 mg product/L group). The 48 h EC50 was determined to be 6.5 mg a.i./L based on nominal concentrations.

 

A study on the acute toxicity of C12 AAPB to Daphnia magna was conducted according to OECD Guideline 202 (April 2004). The daphnids (20/concentration) were exposed to nominal concentrations of 0 (control),11.6, 25.5, 56.2, 124 and 272 mg test item/L, which correspond to the following concentrations of 4.27, 9.39, 20.7, 45.5 and 100 mg solids/ L, for 48 h under semi-static conditions.

The measured concentrations ranged between 112–117% of the nominal concentrations at the middle and highest treatment level throughout the test. No biological effects were observed at the three lower concentrations. The lowest test concentration showed similar ranges from 121–126% of the nominal concentrations but was slightly higher than the required 120% limit of the nominal concentration throughout the test period.

Since the measured test item concentrations were stable during the test period between 98 and 101% of the measured initial concentrations and the overall recovery of the measured concentrations was 118% of the nominal concentrations, the biological results are calculated and reported based on nominal concentrations.

A clear concentration-response relationship was observed. The following EC50 value was determined:

48 h EC50 = 124 mg test item/L, corresponding to 45.6 mg solids/L

 

Two further supporting studies are available:

The acute toxicity of Coco AAPB to Daphnia magna was investigated in a study conducted according to OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) and EU Method C.2 (Acute Toxicity for Daphnia). The daphnids were exposed to nominal concentrations of 0 (control), 0.56, 1.0, 1.8, 3.2, 5.6, and 10.0 mg a.i./L for 48 h under semi-static conditions (renewal of the test solutions after 24 h). The 48 h EC50 was found to be 4.1 mg a.i./L (nominal).

 

The 48-hr-acute toxicity of Coco AAPB to Daphnia magna was investigated in a study conducted according to EU Method C.2 (Acute Toxicity for Daphnia) under static conditions. Daphnids were exposed to control and test chemical at (nominal/measured) concentration of 3.5; 6; 10; 17; 30 mg dry residue/L for 48 hr. Immobilization were observed. The 48 h EC50 was 6.6 mg a.i./L nominal.

 

Marine invertebrates

The acute toxicity of Coco AAPB to the marine copepod  Acartia tonsa  was investigated in a study conducted according to ISO 14669 (1999) Water Quality - Determination of the acute lethal toxicity to marine capepods (Copepoda; Crustacea).  Acartia tonsa  were exposed to nominal Coco AAPB static conditions for 48 h. The 48 h NOEC and LC50 values were determined to be 3.6 mg a. i./L and 7.0 mg a.i./L nominal, respectively. The 48 h LC50 of 7.0 mg a i./L nominal was used for further assessment.

 

Conclusion

For freshwater invertebrates (Daphnia magna) the lowest 48 h EC50 was determined to be 1.9 mg a. i./L (nominal), and for the marine copepod  Acartia tonsa   the 48 h EC50 was 7.0 mg a.i./L (nominal).

 

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 13.

 

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.

 

1. 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.

 

1. 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 invertebrate toxicity data

 

Endpoints	Target substance	Source substances
	C8-18 AAPB	C8-18 and C18 unsatd. AAPB	C12 AAPB
Short-term toxicity to aquatic invertebrates	key_Short-term toxicity to aquatic invertebrates: 97862-59-4_9.1.1_THG_1991c_OECD 202   key study   OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) Daphnia magna, static, freshwater   48 h EC50 = 6.5 mg/L act. ingr. (nominal) based on: mobility (95% C.I.: 4.8 - 8.4 mg a.i./L) 24 h EC50 > 30 mg/L act. ingr. (nominal) based on: mobility   Reliability: 1 (reliable without restriction), GLP	key.Short-term toxicity to aquatic invertebrates: 61789-40-0_9.1.1_KAO Corporation_1992_OECD 202   key study   OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) (part I, 24h EC50 Acute Immobilization Test (1984)) Daphnia magna, static, freshwater   48 h EC50 ca. 1.9 mg/L act. ingr. (nominal) based on: mobility (95% CL: 1.37 - 2.69 mg a.i./L)   Reliability: 2 (reliable with restrictions), GLP	key_Short-term toxicity to aquatic invertebrates: 4292-10-8_9.1.1_Solvay_2020_OECD202   key study   OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test) Daphnia magna, semi-static, freshwater   48 h EC50 = 124 mg/L as test material, corresponding to 45.6 mg solids/L   Reliability: 1 (reliable without restriction), GLP
		key (marine).Short-term toxicity to aquatic invertebrates.61789-40-0_9.1.1__Marine_48h_AcartiaTonsa_Rhodia_Mirataine-BET-C37_ISO_2008   key study   ISO 14669 (1999) Water Quality - Determination of the acute lethal toxicity to marine copepods (Copepoda; Crustacea) Acartia tonsa, static, saltwater 48 h NOEC = 10 mg/L test mat. (nominal) based on: mobility (95% c.i.: 14.85-23.01 mg/L) 48 h NOEC = 3.6 mg/L act. ingr. (nominal) based on: mobility (95% c.i.: 5.4 - 8.3 mg a.i./L)   48 h LC50 = 19.38 mg/L test mat. (nominal) based on: mobility (95% c.i.: 14.85-23.01 mg/L) 48 h LC50 = 7 mg/L  act. ingr. (nominal) based on: mobility (95% c.i.: 5.4 - 8.3 mg a.i./L)   Reliability: 1 (reliable without restriction), GLP
		sup.Short-term toxicity to aquatic invertebrates.61789-40-0_9.1.1_Acute_Daphnia_Unilever_A17-AT-L17-01 supporting study OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)   Daphnia magna, semi-static, freshwater   48 h EC50 = 4.1 mg/L act. ingr. (nominal) based on: mobility (95% c.i.: 3.3-5.3 mg a.i./L)   Reliability: 2 (reliable with restrictions), GLP
		sup.Short-term toxicity to aquatic invertebrates: 61789-40-0_9.1.1_Hüls_1996_EEC 92-69 C2   supporting study   EU Method C.2 (Acute Toxicity for Daphnia) Daphnia magna, static, freshwater   48 h EC0 < 3 mg/L act. ingr. (nominal) based on: mobility 48 h EC100 > 25 mg/L act. ingr. (nominal) based on: mobility 48 h EC50 = 6.6 mg/L act. ingr. (nominal) based on: mobility (95% c.i. 4 - 11 mg/L)   Reliability: 2 (reliable with restrictions), GLP

 

The 48 h EC50 values of C8-18 AAPB and C8-18, C18 unsatd. AAPB to  Daphnia magna were in the range of 1.9 – 6.6 mg a.i./L. The sensitivity of the marine invertebrate Acartia tonsa  was in the same order of magnitude (48 h EC50 = 7 mg a.i./L)

 

 

Quality of the experimental data of the analogues:

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

The studies with Daphnia magna were conducted according to OECD Guideline 202 or EU Method C.2 and were reliable or reliable with restrictions (RL1-2, GLP).

A further study with the saltwater organism Acartia tonsa was conducted according to ISO 14669 (RL1, GLP)

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 substances as presented above and in more detail in the general justification for read across, it can be concluded that the available data from the source substance C8-18 and C18 unsatd. AAPB are also valid for the target substance C8-18 AAPB.

 

For freshwater invertebrates (Daphnia magna) the lowest 48 h EC50 was determined to be 1.9 mg a. i./L (nominal), and for the marine copepod  Acartia tonsa  the 48 h EC50 was 7.0 mg a.i./L (nominal).