3-C12-18-(even numbered)-alkylamido-N,N-dimethylpropan-1-amino oxide

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:

supporting study

Study period:

2012-12-12 - 2012-12-14

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Name of test material (as cited in study report): Amides, C12-18 (even numbered), N-[3(dimethylamino)propyl], N'-oxides
- identifiers: EC 268-938-5, CAS 68155-09-9
- Molecular formula (if other than submission substance): not stated
- Physical state: nearly colourless (pale yellowish), clear liquid
- Analytical purity: UVCB with 35.8% active ingredient in water
- Lot/batch no.: ST02496002
- Expiration date of the lot/batch: April 2014
- Stability under test conditions: unknown in aqueous solution
- Storage condition of test material: keep in closed containers at room temperature
- Density: 0.908 kg/m³ (20°C)
- Solubility: 1.05g/L (critical micelle concentration 0.941 g/L)
- Boiling point: 126.4 °C

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Pooled samples of the test media at the beginning and end of each renewal period (0 and 24 h; 24 and 48 h) were analysed
- Sample storage conditions before analysis: The samples were analysed immediately after sampling by liquid chromatography (HPLC)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Straus
- Source: The stock culture was obtained from the German Federal Environment Agency, Institute for Water, Soil, and Air Hygiene (Berlin).
- Age at study initiation (mean and range, SD): aged less than 24 hours at the start of the test
- Method of breeding: Specimens used in the test were bred in the laboratory at Fraunhofer IME.
- Feeding during test: no
- Food type: fed daily with an algal suspension (Desmodesmus subspicatus containing Artemio Fluid JBL).
- Frequency: daily

ACCLIMATION
- Acclimation period: To maintain the stock culture, batches of 30 - 50 adult daphnids were held at room temperature in approximately 1.8 L of dilution water for one week
- Acclimation conditions (same as test or not):
- Type and amount of food:
- Feeding frequency:
- Health during acclimation (any mortality observed):

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

1.2 mmol/L

Test temperature:

19.7± 1°C

pH:

at start: 8.39-8.53
at end: 8.20-8.35

Dissolved oxygen:

at start: 8.2 - 8.6 mg/L
at end: 7.7-8.6 mg/L

Salinity:

not stated

Nominal and measured concentrations:

Range finder test: nominal concentrations: control, 1, 10, and 100 mg/L (corresponding to 0.36, 3.6, and 35.8 mg a.i./L).
Definitive test: the nominal concentrations of the test item used were 6.25, 12.5, 25.0, 50.0, and 100 mg/L (corresponding to 2.2, 4.5, 9.0, 17.9, and 35.8 mg a.i./L). The concentrations were spaced by a factor of 2 and agreed upon with the monitor.

Details on test conditions:

TEST SYSTEM
- Test vessel: 60 mL glass beakers
- No. of organisms per vessel: 4
- No. of vessels per concentration (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Schmallenberg district water production plants, mostly fed by small springs and percolation. The purification process occurs on site at Fraunhofer IME and includes filtration with activated charcoal, passage through a lime-stone column, and aeration to the point of oxygen saturation

OTHER TEST CONDITIONS
- Photoperiod: The daphnids were maintained under a light/dark cycle of 16/8 hours.
- Light intensity:light intensity did not exceed 1125 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): Mortality and clinical signs (sub-lethal and abnormal behavior) were recorded at 3 h, 24 h, 48 h, 72 h and 96 h after fish addition.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations: The test included one control tank and five tanks with nominal test concentrations of 0.625, 1.25, 2.5, 5.0 and 10 mg test item/L. Analysis of test media revealed concentrations of test item between 96 and 106 % of nominal for fresh, and between 96 and 108 % of nominal for the aged test media. Since one measurement was found to be > 120 % of nominal, the geometric mean was used for the final determination of effect data. The geometric mean measured concentrations were calculated to be 0.663, 1.32, 2.60, 5.08 and 9.59 mg test item/L.
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

K2Cr2O7

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

19.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Results with reference substance (positive control):

EC50: The latest 24h determined EC50 was 1.03 mg/L (April 2012).

Reported statistics and error estimates:

The percent immobilisation (test endpoint) of each test concentration will be compared with a control consisting of dilution water only. If the analyses demonstrate that the deviation from the nominal concentration is above 20 %, all effect data will be based on the measured concentrations. If possible, a NOEC/LOEC is calculated by using ANOVA followed by Dunnett’s or Williams’ tests or an appropriate non-parametric test. When the test results show a concentration-response relationship, the data is analysed by regression to determine the EC50/20/10 including the 95 % confidence interval using Probit-analysis assuming log-normal distribution of the values. The computer software ToxRat® Professional v.210 will be used for statistical evaluations.

Validity criteria fulfilled:

yes

Conclusions:

At 24 h, 5% immobilisation effect was observed in the 100 mg/L test concentration. After 48 h, the concentration dependent effect was more apparent. Immobilisation was 20 and 100% in the 50.0 and 100 mg/L test concentrations resulting in a NOEC of 50.0 mg/L corresponding to 17.9 mg a.s./L. The EC50 was 55.5 mg/L, corresponding to 19.9 mg a.i./L.
No immobility was observed in the control. There were no signs of disease or stress, e.g. discolouration or abnormal behaviour, observed in any replicate. All surviving specimens appeared normal.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1996-09 - 1996-11

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)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Name of test substance: Aminoxid WS 35
- identifiers: EC 268-938-5, CAS 68155-09-9
- Chemical characterisation: Fatty acid-aminoalkyldimethylaminoxid - aqueous solution with 35% active matter
- Batch no.: 571003
- Storage conditions: room temperature
- Expiry date: April 1997
- Chemical stability in water: stable
- Stability under test conditions: supporting analysis study was performed

Analytical monitoring:

yes

Details on sampling:

- Concentrations: screening was performed with the following concentrations: 10,000; 1000; 100; 10 and 1 mg/L resp.
- Sample storage conditions before analysis: sample to be analyzed were stored deep frozen and protected from light until analysis was performed.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: STRAUS
- Source: derived from LWA Dusseldorf, the cultivation was performed according to SOP S 001 of Institut Fresenius Taunusstein
- Age at study initiation (mean and range, SD): >6h and < 24h
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Hardness:

not stated

Test temperature:

20.5 +- 0.5 °C

pH:

start: 8.41 - 8.68
end: 7.81 - 7.89

Dissolved oxygen:

start: 7.9 - 8.0 mg/l O2
end: 7.9 - 7.8 - 8.2 mg/l O2

Salinity:

not stated

Nominal and measured concentrations:

Control, 5, 10, 20, 40, 80, 100, 200 mg a.i./l (nominal)

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: vessels, containing 50 ml of test solution
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates):8
- No. of vessels per control (replicates): 8


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water was prepared according to OECD 202 Annex 3 (ISO water)

OTHER TEST CONDITIONS
- Photoperiod: 16h light / 8h dark


EFFECT PARAMETERS MEASURED:
- immobilisation

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study
- Test concentrations: 10,000; 1,000; 100; 10 and 1 mg/l
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

16 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: immobilisation

Remarks on result:

other: NA

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Reported statistics and error estimates:

Percentage of affected daphnia after 24 and 48h was plotted on probit paper and the value fot EC50 was taken from the graph.

highest concentration causing no mortality after 48h: 10 mg test substance/l

lowest concentration causing 100% mortality after 48h: 80 mg test substance/l

Concentration [mg test substance/l]	Immobilised Daphnia after 24h [%]	Immobilised Daphnia after 48h [%]
control	0	0
200	100	100
100	70	100
80	50	100
40	10	40
20	0	5
10	0	0
5	0	0

Results of analytical monitoring:

The concentrations were within 100% +- 20% of nominal concentration at test start and test termination. Therefore the effect values are reported as nominal concentrations.

Validity criteria fulfilled:

yes

Conclusions:

In an acute toxicity study with Daphnia magna according to OECD 202 a 48h-EC50 of 46 mg test substance/l based on nominal concentrations was calculated. The test substance Aminoxide WS 35 is of moderate toxicity to daphnids.

Executive summary:

The 48 h-acute toxicity of AMINOXIDE WS 35 to Daphnia magna was studied under static conditions according to OECD guideline 202. Daphnids were exposed to control, and test chemical at (nominal) concentrations of 5, 10, 20, 40, 80, 100, 200 mg test substance/l for 48 hr. Immobilization was observed daily. The 48 h-EC50 was 46 mg test substance/l.

This study is classified as reliable without restriction (Guideline study (OECD 202), validity criteria of the guideline fulfilled) and satisfies the guideline requirements for an acute toxicity study with freshwater invertebrates.

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1999-12-01 - 1999-12-03

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Name (as cited in study report): N-(3-N,N-dimetylainooxidetrimetylene)alkylamide
- Code name: NINOX HCDO
- Purity: 92%
- Lot/batch no.: 876 TK Sample IX
- Expiration date of the lot/batch: not stated
- Storage conditions of test material: dark place at room temperature

Analytical monitoring:

yes

Details on sampling:

- Concentrations: water samples were taken from the control and the 0.1, 0.32, 1.0, 3.2 and 10 mg a.i./L test groups (replicates R1-R2 pooled) at 0 and 48 h for quantitative analysis
- Sampling method: duplicate samples and samples at the 0.18, 0.56, 1.8 and 5.6 mg a.i./L test group were taken and stored frozen for analysis
- Sample storage conditions before analysis:

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: not stated
- Source: maintained in a laboratory culture originating from a strain supplied by the Institut National de recherche Chimique Appliqueé (IRCHA), France


ACCLIMATION
- Acclimation period: 24 h
- Acclimation conditions (same as test or not):
- Type and amount of food: suspension of mixed algae
- Feeding frequency: daily
- Health during acclimation (any mortality observed): no

Test type:

static

Water media type:

freshwater

Total exposure duration:

48 h

Hardness:

250 mg/L CaCO3

Test temperature:

21°C

pH:

7.8 ± 0.2

Dissolved oxygen:

reconstituted water was aerated until the DO concentration was approx. air-saturation value

Salinity:

not stated

Nominal and measured concentrations:

range-finder: nominal concentrations of 0.1, 1.0, 10 and 100 mg a.i./L
definitive study: 0.1, 0.18, 0.32, 0.56, 1.0, 1.8, 3.2, 5.6, 10 mg a.i./L

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 ml glass jars containing 250 ml of test solutions
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume:
- Aeration:
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate): no
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2

OTHER TEST CONDITIONS
- Photoperiod: ligthing cycle was controlled to give a 16 h light and 8 h darkness cycle with 20 min. dawn and dusk transition periods

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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations: 0.1, 1.0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.96 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: 0.7-1.2

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.32 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

act. ingr.

Basis for effect:

mobility

Reported statistics and error estimates:

analysis of the immobilisation data was done by probit method of Finney (1971) at 24h and 48 h

Validity criteria fulfilled:

yes

Conclusions:

The 48 h EC50 based on nominal test concentrations was 0.96 mg a.i./L with 95% confidence limits of 0.7-1.2 mg a.i./L.

Executive summary:

Analysis of the test preparations at 0 and 48 hrs showed the measured test concentrations to be in the range of 80% to 100% of nominal with the exception of the 0.1 mg a.i./L test concentration at 48 hrs which was found to be 76% of nominal. Given that this test concentration was below the NOEC and hence had no bearing on the overall results of the study, it was considered justifiable to base the results on nominal test concentrations only.

Description of key information

One study (Wetton and Mullee, 2000) is considered as the key study for endpoint coverage. The study investigates the acute toxicity of the substance to Daphnia magna according to OECD guideline 202, under static conditions in freshwater and with an exposure period of 48 hour. The 48h-EC50 was determined to be 0.96 mg a.i./L . The study is given a Klimisch score of 1 and was conducted under GLP.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

0.96 mg/L

Additional information

Three studies investigating the acute toxicity of the substance to aquatic invertebrates according to OECD guideline 202 are available. All three studies are given a Klimisch score of 1 and were conducted under GLP.

 

The study of Wetton and Mullee (2000) is selected as key study for endpoint coverage. The acute toxicity of the test substance to the freshwater invertebrate Daphnia magna was investigated and gave a 48h-EC50 of 0.96 mg a.i./L.

 

The studies of Kamp (2013) and Schlechtriem (2013) are considered as supporting studies for endpoint coverage. They determined 48h-EC50 values of 16 mg a.i./L and 19.9 mg a.i./L respectively.