reaction product of wheat aminoacids and lauryl chloride

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29.April.2010-21.July.2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD guideline, GLP study

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

Deviations:

yes

Remarks:

One deviation was observed during test n°1: the algae preculture was run four days prior to the test instead of three days. As a consequence, its concentration was prepared at 5.10^3 instead of 1.10^4 cell /ml. This deviation has any influence on results.

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:

The algae cultures are stored in the laboratory on a solid medium (LC/agar Petri plates) and in a liquid medium with regular sub-culturing (every week).
Four days prior to the test, a preculture of algae was run in the same conditions than the test, in order to have a log phase algae culture for the inoculation. Before inoculation, this preculture was checked under a microscope with a heamocytometer in order to control the algae morphology and to measure the cell number.

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 algae medium). For that reason, test solutions were prepared from a 100 mg/l stock solution of the test item previously diluted in algae dilution water.

Test organisms (species):

Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Raphidocellis subcapitata
- Strain: CCAP 278.4

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Post exposure observation period:

Not applicable

Test temperature:

Test n°1: 22.5 to 23.5 °C
Test n°2: 22.4 to 22.7 °C

pH:

7.9

Nominal and measured concentrations:

In order to check, the initial concentrations and maintenance of the exposure concentrations during the ecotoxicological testing, a test item analysis was needed and the total organic carbon was chosen as a tracer. The test item was measured at the 10 and 100 mg/l test item concentrations and in dilution water controls at the beginning and at the end of the test.

Details on test conditions:

Light intensity:
Test n°1: 5946 lux
Test n°2: 6271 lux

Culturing apparatus: Cooling incubator Infors Multitron

Dilution water batch number:
test n°1: 7/27041001
test n°2: 7/01071001

Reference substance (positive control):

yes

Remarks:

The last algae test (February 2010) performed on the reference item K2Cr2O7 150210, gives an ErC50 (0-72h) = 1.16 mg/l (1.0-1.3), indicating a correct sensitivity of the algae which conforms well with the historical results of the laboratory.

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

136.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

1.33 g/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Results with reference substance (positive control):

ErC50 (0-72h) = 1.16 mg/l

Test item analyses

Results obtained with biotic samples (with algae)
Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference	algae biomass at 72h (in algae cell/ml)
Dilution water control (6 samples)	7.0	7.3	0.0	21.5	5.2	16.3	+16.3 mg/l	97.8*10^4
10.0 mg/l (3 samples)	9.8	7.2	2.6	17.5	6.2	11.3	+8.7 mg/l	84.6*10^4

total organic carbon concentrations measured in mg/l at the start and the end of the test for biotic samples during test 1.

Results obtained with biotic samples (with algae)

Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference	algae biomass at 72h (in algae cell/ml)
Dilution water control (6 samples)	6.8	6.8	0.0	27.2	4.3	22.9	+22.9 mg/l	181.2*10^4
10.0 mg/l (3 samples)	9.6	6.7	2.9	28.3	4.4	23.9	+21 mg/l	175.3*10^4
100.0 mg/l (3 samples)	35.7	6.7	29.0	35.0	7.0	28.0	-1.0 mg/l	55.3*10^4

total organic carbon concentrations measured in mg/l at the start and the end of the test for biotic samples during test 2.

Results obtained with abiotic samples (without algae)
Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference
Dilution water control (6 samples)	7.0	7.5	0.0	7.0	7.0	0.0	0 mg/l
10.0 mg/l (3 samples)	9.7	7.2	2.5	7.8	7.1	0.7	-1.8 mg/l

total organic carbon concentrations measured in mg/l at the start and the end of the test for abiotic samples during test n°1.

Results obtained with abiotic samples (without algae)
Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference
Dilution water control (6 samples)	6.8	6.8	F/b dated  0.0	6.6	6.7	0.0	0 mg/l
10.0 mg/l (3 samples)	9.6	6.7	2.9	6.6	6.7	0.0	-2.9 mg/l
100.0 mg/l (3 samples)	35.7	6.7	29.0	37.1	6.7	30.4	+1.4 mg/l

total organic carbon concentrations measured in mg/l at the start and the end of the test for abiotic samples during test n°2.

Validity criteria fulfilled:

yes

Conclusions:

Test 1: ErC50 = 136.6 mg/l (extrapolated value expressed in nominal concentration)
Test 2: ErC50 = 1.33 g/l (extrapolated value expressed in nominal concentration)
The No Observable Effect Concentration (NOEC) and Low Observable Effect Concentration (LOEC) were identical during both tests and have been determined:
NOEC= 10.0 mg/l
LOEC = 100.0 mg/l

Executive summary:

Toxicity of the test item "LCE1001" was performed on the freshwater unicellular green algae Pseudokirchneriella subcapitata according to the OECD 201 guideline: two screening tests were performed (see amendment n°1). The results obtained show, at the highest tested concentration of 100 mg/l (nominal concentration), a toxicity of the test item with an inhibition of the average specific growth rate in comparison to the control equal to 50.0% during test 1, and 22.8% during the second test.

ErC50 (0 -72h) were superior to this concentration and were determined according to the OECD 201 (March 2006):

Test 1: ErC50 = 136.6 mg/l (extrapolated value expressed in nominal concentration)

Test 2: ErC50 = 1.33 g/l (extrapolated value expressed in nominal concentration)

The No Observable Effect Concentration (NOEC) and Low Observable Effect Concentration (LOEC) were identical during both tests and have been determined:

NOEC= 10.0 mg/l

LOEC = 100.0 mg/l

In order to check the initial concentrations and maintenance 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 end of the test.

Note: for test n°1, only the concentration at 10 mg/l was analysed.

Moreover, to verify the possible effects of the presence of biological organism on the behaviour of the test item, additional abiotic flasks without algae (6 for control, 3 for the 10 and 3 for the 100 mg/l) were put in the test and analysed at the beginning and the end of the test. Samples used for these analyses were directly collected from the corresponding flask at concentrations selected.

Results obtained during both tests performed are presented as follows:

Results obtained with biotic samples (with algae)
Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference	algae biomass at 72h (in algae cell/ml)
Dilution water control (6 samples)	7.0	7.3	0.0	21.5	5.2	16.3	+16.3 mg/l	97.8*10^4
10.0 mg/l (3 samples)	9.8	7.2	2.6	17.5	6.2	11.3	+8.7 mg/l	84.6*10^4

total organic carbon concentrations measured in mg/l at the start and the end of the test for biotic samples during test 1.

       

Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference	algae biomass at 72h (in algae cell/ml)
Dilution water control (6 samples)	6.8	6.8	0.0	27.2	4.3	22.9	+22.9 mg/l	181.2*10^4
10.0 mg/l (3 samples)	9.6	6.7	2.9	28.3	4.4	23.9	+21 mg/l	175.3*10^4
100.0 mg/l (3 samples)	35.7	6.7	29.0	35.0	7.0	28.0	-1.0 mg/l	55.3*10^4

total organic carbon concentrations measured in mg/l at the start and the end of the test for biotic samples during test 2.

Results obtained with abiotic samples (without algae)
Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference
Dilution water control (6 samples)	7.0	7.5	0.0	7.0	7.0	0.0	0 mg/l
10.0 mg/l (3 samples)	9.7	7.2	2.5	7.8	7.1	0.7	-1.8 mg/l

total organic carbon concentrations measured in mg/l at the start and the end of the test for abiotic samples during test n°1.

Results obtained with abiotic samples (without algae)
Nominal test item concentrations	Concentrations at T0			Concentrations at T72h
	TC	IC	TOC	TC	IC	TOC	TOC difference
Dilution water control (6 samples)	6.8	6.8	F/b dated  0.0	6.6	6.7	0.0	0 mg/l
10.0 mg/l (3 samples)	9.6	6.7	2.9	6.6	6.7	0.0	-2.9 mg/l
100.0 mg/l (3 samples)	35.7	6.7	29.0	37.1	6.7	30.4	+1.4 mg/l

total organic carbon concentrations measured in mg/l at the start and the end of the test for abiotic samples during test n°2.

The study was performed according to the study plan n° 09/1169F/b dated 23rd April 2010 and its amendment (see annex 1), and in compliance with Good Laboratory Practice (GLP) requirements.

Description of key information

The No Observable Effect Concentration (NOEC) and Low Observable Effect Concentration (LOEC) were identical during both tests and have been determined:

NOEC= 10.0 mg/l

LOEC = 100.0 mg/l