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Short-term toxicity to aquatic invertebrates

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Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
read-across based on grouping of substances (category approach)
Adequacy of study:
weight of evidence
Justification for type of information:
REPORTING FORMAT FOR THE CATEGORY APPROACH
Registered substance (as identified in Section 1.1 identification) has similar chemical structure as the others members of the Category "Amino Acid Alkyl Amides, Sodium or
Potassium Salts" (the source chemicals).

The category “Amino Acid Alkyl Amides, Sodium or Potassium Salts” has been developed and evaluated to cover the assessment of the intrinsic properties of several similar
substances within the context of REACH Regulation. The Category includes sodium or potassium salts of amino acid alkyl amides, with a carbon chain length range of C6-C16 and an
aminoacidic pool of not selected aminoacids reflecting the natural distributions of aminoacids in various vegetables and cereals.


1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
The Category has been built up considering the common chemical structure of its members. They are UVCB substances and they share the following features:
- they are salts of amino acid alkyl amides
- they are sodium or potassium salts
- the carbon chain length ranges between C6 and C16
- the carbon chain derives from cocoyl or lauroyl fatty acids
- the aminoacidic part of the molecules comes from a pool of aminoacids
- the aminoacids pool reflects the natural distributions of aminoacids in various vegetables and cereals.

Overall, the Category was formed on the hypothesis that substances sharing this common chemical structure can be considered analogues and can share intrinsic properties too. No
trends in properties is expected, except for those physicochemical properties depending on the carbon chain length.

2. CATEGORY APPROACH JUSTIFICATION
These substances share (i) eye damage and (ii) the absence of significant systemic toxicity.
Key result
Remarks on result:
other: No aquatic toxicity are expected.
Remarks:
All results on category members support the lack of aquatic toxicity.
Validity criteria fulfilled:
yes
Conclusions:
A collection of the aquatic toxicity data is available for category members. All results support the lack of aquatic toxicity. Overall, considering (i) aquatic toxicity values (ii) the ready biodegradability, the registered substance is not expected to be toxic.

According to Regulation (EC) n. 1272/2008, the registered substance should not be classified for the aquatic toxicity because the available data are judged as "conclusive but not sufficient for classification".
Executive summary:

The category “Amino Acid Alkyl Amides, Sodium or Potassium Salts” was formed on the hypothesis that substances sharing common chemical structure can be considered analogues and can share intrinsic properties too.

Based on this assumption, this property may be assessed using available data  for the category members.

The category has been assessed for members characterized by:

- sodium or potassium salts

- the carbon chain length ranges between C6 and C16

- the aminoacids pool reflects the natural distributions of aminoacids in various vegetables and cereals.

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2004-03-12 to 2004-06-09
Reliability:
2 (reliable with restrictions)
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
no
Details on sampling:
Not applicable
Vehicle:
no
Details on test solutions:
The mother solution preparation is carried out at a concentration a little bit higher than 100 mg/L.
Dilutions are made in order to obtain concentrations in a geometrical series; that permit to calculate the EC50-48 hours; the concentrations are; 0.6; 1.1; 5.6; 11; 56 and 111 mg/L.
The test item solution has to be prepared immediately before the introduction of Daphnia's.
Test organisms (species):
Daphnia magna
Details on test organisms:
Daphnia magna Straus aged more than 6 hours and less than 24 hours at the beginning of the test.
Source: U.G. Gent University_laboratory Prof.Persoone. In incubation in the laboratory
Alimentation: Cultivation of Selenastrum capricornutum algae
Test type:
not specified
Water media type:
not specified
Limit test:
no
Total exposure duration:
48 h
Post exposure observation period:
no post exposure observation period
Hardness:
250 mg/l (in CaCO3)
Test temperature:
21°C ± 1°C.
pH:
pH 7.6 (± 0.2)
Dissolved oxygen:
No data
Salinity:
No data
Nominal and measured concentrations:
Nominal concentrations: 0.6; 1.1; 5.6; 11; 56 and 111 mg/L.
Details on test conditions:
20 organisms by test concentration (2 parts of 10).
Reference substance (positive control):
no
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
ca. 45 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mobility
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
no data
Validity criteria fulfilled:
not specified
Conclusions:
After a 48 hours incubation, the immobilisation of 50% Daphnia’s magna appears for a 45 mg/l concentration.
EC50 48h Daphnia magna: 45 mg/l
Executive summary:

Determination, in the conditions defined in this norm, of the initial concentration (it means the concentration at the beginning of the test) that, in 24 and 48 hours, immobilises 50 % of the Daphnia’s put in experimentation.

This concentration called effective initial inhibitive concentration is designed by EC50-24/48 h.

After a 48 hours incubation, the immobilisation of 50% Daphnia’s magna appears for a 45 mg/l concentration.

EC50 48h Daphnia magna: 45 mg/l

Endpoint:
short-term toxicity to aquatic invertebrates
Type of information:
experimental study
Adequacy of study:
key study
Study period:
17.May.2010-20.May.2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD guideline, GLP study
Qualifier:
according to guideline
Guideline:
OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)
GLP compliance:
yes (incl. QA statement)
Specific details on test material used for the study:
Test item:
Name: LCE1001
FCBA reference: 09/1169F
Batch number: 0915600014
Manufacturing date: June 5th 2009
Composition: amino acid N-coco acyl: 90% / acid lauric 10%
Manufacturer: SEPPIC
Test item appearance: liquid
Test item quality: 50 ml
Packaging: plastic bottle
Storage conditions: room temperature
Stability, deadline for use: 5th June 2011
Reception date of the test item: 11th February 2010

Reference item description:
Name: potassium dichromate
Supplier: accros organics
Batch number: A0242511
CAS number: 7778-50-9
Purity > 99%

Analytical monitoring:
yes
Details on sampling:
In order to check the initial concentrations and maintenance of the exposure concentrations during the ecotoxicological testing, a tes item analysis was needed and the total organic carbon was chosen as tracer: the test item was measured at the 10 and 100 mg/l test item concentrations and in both dilution water controls at the start and end of the test. Moreover, to verify the possible effects of the presence of biological organism on the bahaviour of the test item, 12 abiotic test containers (6 for the control, 3 for the 10 mg/l concentration and 3 for the 100 mg/l concentration) were put in the test and analysed at the beginning and the end of the test.
These analysis were performed according to the document "Fiche de Méthode FCBA CHIMIE n° 209 version 1: Dosage de formulation LCE10001 de SEPPIC par mesure du Carbone Organique Total dans les milieux écotox algues et daphnies" (see final report n° 402/09/1169F/a-e in annex 3). This document describes the full details for the analysis of the test item through Total Organic Carbon analysis in samples from Ecotoxicology testing (preparation and analytical conditions, statistical results).
Samples were directly collected from the corresponding tests containers at concentrations selected, and were analysed according to the recommendations specified in the analytical protocol cited above.
Vehicle:
yes
Details on test solutions:
Test item preparation:
Test item is considered as soluble by the sponsor, in a weak saline aqueous medium (corresponding to the daphnids medium). For that reason, test solutions were prepared from a 100 mg/l stock solution of the test item previously diluted in daphnids dilution water.
Protocol of the test:
The protocol consisted in preparing the three concentrations (1, 10 and 100 mg/l) in daphnids dilution water and in exposing daphnids to these solutions. Two kinds of controls were used in this study: the dilution water control containing daphnids without test item and a abiotic dilution water control without daphnids or test item. This one was used in order to check the contribution of Organic Carbon by daphnids in the medium at the end of the test.
The test was carried out under semi-static conditions, which means that all the test solutions were renewed at 24h. For each concentration, 4 testing containers were prepared, each containing 10 ml of test solution and 5 daphnids. For each both controls, 6 tests containers were prepared. Tests containers were covered in order to avoid evaporation and dust deposition.

Test organisms (species):
Daphnia magna
Details on test organisms:
TEST ORGANISM
- Common name: Daphnia Magna Straus
- Strain/clone: clone number 5, from at least the third generation (i.e. the third brood) obtained with a non-cyclic parthenogenesis reproduction.
- Feeding during test: daphnids were not fed during the test period
The daphnids breeding is carried out in closed bottles placed in a climatic chamber free from any toxic vapour.
Daphnids used in this experimentation were previously filtered in order to obtain animals from a size between 560 and 800 micrometer (corresponding to daphnids less than 24 hours).
Test type:
semi-static
Water media type:
saltwater
Limit test:
no
Total exposure duration:
48 h
Remarks on exposure duration:
The test duration was 24 h with an extension of 48 h
Post exposure observation period:
Not applicable
Hardness:
270 mg/l of CaCO3
Test temperature:
18.9 - 20 °C
pH:
7.6
Dissolved oxygen:
9.1 mg/l
Salinity:
Tes solutions theoritical concentrations:
CaCl2, 2 H2O 297 mg/l
MgCl2, 6 H20 167 mg/l
NaHCO3 200 mg/l
K2SO4 26 mg/l
Reference substance (positive control):
yes
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Key result
Duration:
24 h
Dose descriptor:
NOEC
Effect conc.:
ca. 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Key result
Duration:
48 h
Dose descriptor:
NOEC
Effect conc.:
ca. 10 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
other: immobilisation
Details on results:
No sign of stress was observed on the mobile daphnids after 48 hours of exposition.
Results with reference substance (positive control):
The last toxic response obtained on the reference item K2Cr2O7 030510 (03rd May 2010) gives an EC50-24h of 0.98 mg/l (0.89 - 1.08) indicating that the daphnids sensitivity is correct (within the accepted interval of 0.6 - 2.1 mg/l fixed by the international standard ISO 6341 May 1996) and in accordance with the historical data obtained by the laboratory.

Results on the test item:

Daphnids immobilisation

Raw data (daphnids immobilisation in cumulative number for each observation time, 24 and 48 hours) are presented in table hereafter:

 Nominal test item concentrations in mg/l  Number of daphnids exposed at the start   Immobilised daphnids at 24h  Immobilised daphnids at 48h 
Dilution water control   20  0  0
 Abiotic control  0  0  0
 1.0  20  0  0
 10.0  20  0  0
 100.0  20  0  2
Validity criteria fulfilled:
yes
Conclusions:
The results obtained on the ecotoxicological endpoint "immobilisation" give an EC50-48h superior to the highest tested concentration (i.e 100 mg/l), value expressed in mg/l of the nominal item concentration.
This result, expressed in mg/l of the nominal test item concentration, shows that no toxicity was observed in the concentrations selected in the test on the micro crustacean Daphnia magna. No sign of stress was observed on the daphnids during the 48 hours of the experiment.
Executive summary:

The acute immobilisation screening test of the micro crustacean Daphnia magna was performed on the test item "LCE10001" with semi-static conditions. The results obtained on the ecotoxicological endpoint "immobilisation" give an EC50-48h superior to the highest tested concentration (i.e 100 mg/l), value expressed in mg/l of the nominal item concentration.

This result, expressed in mg/l of the nominal test item concentration, shows that no toxicity was observed in the concentrations selected in he test on the micro crustacean Daphnia magna. No sign of stress was observed on the daphnids during the 48 hours of the experiment.

In order to check the initial concentrations and maintenant of the exposure concentrations during the ecotoxicological testing, a test item analysis was needed and the total organic carbon was chosen as tracer; the test item was measured at the 10 and 100 mg/l test item concentrations and in dilution water controls at the start and at the end of the test. Moreover, to verify the possible effects of the presence of biological organism on the behaviour of the test item, 12 abiotic test containers (6 for the control, 3 for the 10 mg/l concentration and 3 for the 100 mg/l concentration) were put in the test and analysed at the beginning and at the end of the test.

Samples used for these analysis were directly collected from the coresponding flasks at concentrations selected.

Results are presented as follows:

 Nominal test item concentrations        TOC concentrations (samples with daphnids)        TOC concentrations (abiotic samples, without daphnids)
   at 0h  at 24h*  difference  at 0h  at 24h*  difference
 Dilution water control (6 samples)  0.3  0.4  +0.1 mg/l  0.3  0.2  -0.1 mg/l
 10.0 mg/l (3 samples)  3.8  3.8  0 mg/l  3.8  4.7  +0.9 mg/l
 100.0 mg/l (3 samples)  29.5  31.9  +2.4 mg/l  29.5 31.1   +1.6 mg/l

* Because of semi-static conditions, samples were aged of 24h instead of 48h.

Because the results obtained in both controls are almost identically, the presence of daphnids don't affect the TOC concentrations.

The TOC increase observed in the abiotic 10 mg/l concentration is not significant because of the sensitivity of the analysis method. Thus, all the test item concentrations measured have been satisfactorily maintained within +/- 20 per cent of the nominal concentration during the test.

This study was performed according to the study n°09/1169F/c dated 23rd April 2010 and in compliance with Good Laboratory Practice (GLP) requirements.

Description of key information

Category members (Amino Acid Alkyl Amides, Sodium or Potassium Salts) were not found to be significant toxic to neither aquatic invertebrates nor aquatic algae.

Key value for chemical safety assessment

Additional information

A collection of the aquatic toxicity data are available for category members. All found values are listed below. 

EC number

Short-term toxicity to aquatic invertebrates

S1 - EC: 918-984-3

EC5048 h Daphnia magna: 45 mg/l

S2 - EC: 927-837-2

The results obtained on the ecotoxicological endpoint "immobilisation" give an EC5048h superior to the highest tested concentration (i.e 100 mg/l), value expressed in mg/l of the nominal item concentration.

No sign of stress was observed on the daphnids during the 48 hours of the experiment.

 

Overall, considering (i) aquatic toxicity values (ii) the ready biodegradability, the registered substance is not expected to be toxic.

According to Regulation (EC) n. 1272/2008, the registered substance should not be classified for the aquatic toxicity because the available data are judged as "conclusive but not sufficient for classification".