reaction product of wheat aminoacids and lauryl chloride

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

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

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

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

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

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.