N-(1-oxooctyl)glycine

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

june 12 2008-November 11 2008

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: according to Guideline OECD 202

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

not applicable

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

N-(1-oxooctyl)glycine

EC Number:

238-122-3

EC Name:

N-(1-oxooctyl)glycine

Cas Number:

14246-53-8

Molecular formula:

C10H19NO3

IUPAC Name:

N-(1-oxooctyl)glycine

Details on test material:

- Name of test material (as cited in study report): LCA07012
- Substance type: mono-constituant
- Physical state: solid
- Analytical purity: >97%
- Lot/batch No.: 716300006
- Expiration date of the lot/batch: 11.06.2010
- Stability under test conditions: yes
- Storage condition of test material: room temperature (20 ± 5 °C)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Melting point: 105-110°C
- Vapour pressure: 1.33*10-4Pa
- Water solubility (under test conditions): 1560mg/l
- log Pow: 2.052 ± 0.02
- Stability in water: H2O: < 0.1g/L

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

storage: at room temperature

Test solutions

Vehicle:

no

Details on test solutions:

As solubility was determined with about 1560 mg/L (AB07080601G910), a test solution of an appropriate concentration 100 mg/L in dilution water was prepared. The resulting solution was clear and homogenous

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
Species Daphnia magna
Variety STRAUS
Strain Berlin
Sex female
Age between 0 and 24 hours
Origin Umweltbundesamt Berlin
Arrival of Strain 27. Sep. 2007
Selection of the test system was made following the proposal of the guidelines.


Animal Husbandry
Daphnia magna is bred in the LAUS GmbH throughout the year. The animals are kept for the use in toxicity tests. They multiply by parthenogenesis, thus being genetically identical. The holding is performed similar to the method described in EN ISO 6341, following SOP 115 002 01 („Zucht und Hälterung von Daphnia magna STRAUS“), edition 9, adopted 27. Apr. 2007.
Vessels glass beakers, nominal volume 2 l
Medium M4-Medium (recipe of ELENDT), composition see annex
Food unicellular green algae (Desmodesmus subspicatus)
Medium renewal twice a week
Photo period 16/8 hours, using neon tubes
Temperature 20  2 °C

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

24 h

Post exposure observation period:

The test vessels were left to stand for 48 hours. After 24 and 48 hours, the immobilised daph-nia were counted. Daphnia are considered immobilised if they perform no movements or are only able to move their antennae when the beaker is gently agitated.
The pH, the concentration of dissolved oxygen and the content of the test item in the test ves-sels were measured at the beginning and at the end of the test.
All values were documented in the raw data.

Test conditions

Test temperature:

25.5 – 26.3 oC

pH:

7.0 to 7.8.

Dissolved oxygen:

above 7.8 mg/L

Nominal and measured concentrations:

Concentration nominal :100mg/L
Measured Conc. at 24 and 48 h:103.00 and
113.95 mg/L

Details on test conditions:

Vessels glass beakers, nominal volume 2 l
Medium M4-Medium (recipe of ELENDT), composition see annex
Food unicellular green algae (Desmodesmus subspicatus)
Medium renewal twice a week
Photo period 16/8 hours, using neon tubes
Temperature 20  2 °C

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Effect concentrations 2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Details on results:

Immobilities
In the control, none of the daphnia died or showed any signs of abnormal behaviour through-out the test (see table below). After 24 hours in treatment 100 mg/L, 50 % of the animals were immobilised on the surface of the test solution. After 48 hours, this effect wasn’t observed any more. Only 10 % of the daphnia were immobilise

Results with reference substance (positive control):

Biological Results Reference Item
The estimation of the EC50 was accomplished using the software OriginTM. The calculated values for p resp. χ2 are given in the graph.
The data were evaluated using sigmoidal fit on a linear-logarithmic scale.


The biological results are presented in the following table:
Biological Results Reference Item
PARAMETER Value 95%-confidence interval
NOEC 24 h 0.5 mg/L not determinable
24h EC50i 0.85 mg/L not determinable
24h EC100i 2.0 mg/L not determinable
n. d. = not determinable

Reported statistics and error estimates:

none

Any other information on results incl. tables

 Analytical Determinations

Inadvertently, the samples of t = 0 h were not measured. See discussion (chapter10, page23).

The measured concentrations showed very good correlation with the nominal concentrations. The measured concentration lay between 103 and 114 % of the nominal concentration. The recovery rate after 24 hours was 106%

Therefore, the nominal concentrations were used for the determination of the results.

The measured concentrations of the test item can be found in the following table.

 Measured Concentrations

Time	Treatment (Nominal Concentration)	New Solution (Medium renewal every 24 h)		Old Solution (Medium renewal every 24 h)		Recovery after 24 h (based on Measured Conc.)
		Measured Conc. at 24 and 48 h	% of Nominal Conc.	Measured Conc. at 24 and 48 h	% of Nominal Conc.
hours	mg/L	mg/L	%	mg/L	%	%
0-48	0	< LOQ	--	< LOQ	--	--
24	100	107.54	108	103.00	103	--
48	100	--	--	113.95	114	106

LOQ = Limit of quantification

The increase is attributed to evaporation effects

Biological Results Test Item

The biological results are presented in the following table:

Biological Results Test Item

Parameter	Value	95%-confidence interval
NOEC 24h	100 mg/L	not determinable
24h EC50i	> 100 mg/L	not determinable
24h EC100i	> 100 mg/L	not determinable
NOEC 48h	100 mg/L	not determinable
48h EC50i	> 100 mg/L	not determinable
48h EC100i	> 100 mg/L	not determinable

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

All validity criteria were met. For the estimation of the EC50s of the reference item, the fits showed sufficient statistical correspondence of the data with the dose-response-equation.
The 24h-EC50i of potassium dichromate was determined as 0.85 mg/L, which is within the demanded range of 0.6 – 2.1 mg/L.
The analytical determinations of the test item showed good correlation between nominal and measured concentrations and good recovery rates. Therefore, the determination of the results was based on the nominal concentrations. Inadvertently, the analytical samples of t = 0 h were not measured (vials were dropped and broke during transport to HPLC). As the other deter-minations verify that the test item LCA07012 was stable under the test conditions, this was stated as uncritical.
In the first main study, 50 % of the animals were found immobilised on the surface of the test solution 100 mg/L after 24 hours. After 48 hours, this effect wasn’t observed any more: only 10 % of the daphnia were immobilised. Immobilisation was therefore reversible and regarded as uncritical for the assessment of the risk to the aquatic environment. Nevertheless, the main study was repeated (without analytical determination) in order to validate this result. In the second main study, no immobilisation was observed. The difference in the behaviour of the animals is not unusual for biological test systems. As in both main studies, no irreversible im-mobilisation surpassing 10% (≤ 10% is stated in the guideline as validity criterion for the negative controls) was found, the result of the study is considered as valid.

Executive summary:

In the pre-test, the concentrations 100; 10 and 1 mg/L were tested. Twenty daphnia were exposed to the test item for 48 hours in a static test system. After 24 and 48 hours, the immobilised daphnia were counted.

The first main study was performed as a limit test using the nominal concentration of 100 mg/L in a semi-static test with medium renewal every 24 hours

After 24 hours 50 % of the daphnia were immobilised at the surface of the test solution. After 48 hours none of the animals were immobilised in the control, and 10 % were immobilised in the treatment. In order to check this result (EC50_i> 100 mg/L) the main study was repeated (without analytical determination). In the second main study, no animals were immobilised in the treatment and in the control.

The 24h-EC50_iof potassium dichromate was tested in a current reference test. The value was determined as 0.85 mg/L, lying within the demanded range of 0.6 – 2.1 mg/L.

In the first main study, the content of the test item in the old and the new test solutions was determined using HPLC-determination. The recovery after 48 hours lay between 80 and 120  % of the start concentration, the correlation between nominal and measured concentration was very good in all measurements. Therefore, the determination of the biological results was based on the nominal concentration.

The following results could be determined for the test itemLCA07012(species:Daphnia magna).

 

24h-NOEC = 100 mg/L.
48h-NOEC = 100 mg/L
24h-EC50_i> 100 mg/L
48h-EC50_i> 100 mg/L