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

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

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 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

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:

Sample to be analyzed were stored deep frozen and protected from light until analysis was performed.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: Fischzucht Aquafarm Ryba
- Age at study initiation (mean and range, SD):
- Length at study initiation: 1-3 cm
- Feeding during test: no

Test type:

semi-static

Water media type:

freshwater

Total exposure duration:

96

Remarks on exposure duration:

h

Test temperature:

24 - 25 °C

pH:

7.14 - 8.11

Dissolved oxygen:

oxygen content of > 60% of saturation value

Nominal and measured concentrations:

1.0, 2.0, 4.0, 8.0, 10 mg active matter/l (nominal)
results of analytical monitoring in the two highest concentrations: 4.4, 7.8 mg active matter/l (mean measured concentration)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- fill volume: 3.5 l
- Aeration: during the incubation period (96 h)
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate:


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: drinking tapwater
- test substance was dissolved in water by stirring with a magnetic stirrer at room temperature
- renewal of test solutions after 48 h due to high loss in substance concentrations

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


EFFECT PARAMETERS MEASURED:
- mortality, recorded after 24, 48, 72 and 96 h


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study
- Test concentrations: 0.01; 0.1; 1; 10; 100; 1,000; 10,000 mg a.i./l
- Results used to determine the conditions for the definitive study: yes

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.06 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

5.9 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

highest concentration with 0% effect: 4.4 mg test substance/l (measured)
lowest concentration with 100% effect: 7.8 mg test substance/l (measured)
The LC50 values was performed at the geometric mean of the highest concentration (LC0) showing no effect and the lowestconcentration with total effect (LC100).

Sublethal observations / clinical signs:

Results of analytical monitoring

Nominal conc. [mg/l]	Measured conc. [mg/l]	% concentration of Amineoxide XS35 found	Sampling time [hs]	Exporure time [hs]
8	6.2 4.7 /5.3 < 0.8 8.6 < 0.8	77.4 mean: 62.7 < 10 107 < 10	0 24 48 48 96	0 24 48 0 48
10	8.6 7.0	85.6 69.8	0 24	0 24

Nominal conc. [mg/l]	Calculated mean recovery of 48h exposure time	Calculated mean recovery of 48h exposure time	Calculated mean recovery of 24h exposure time	Calculated mean recovery of 24h exposure time
8	4.0 mg/l 4.7 mg/l total mean: 4.4 mg/l (n02)	50.0% (n03) 58.5% (n02) total mean: 54.3% (n=2)
10			7.8 mg/l	77.7% (n=2)

RESULTS OF THE TOXICITY TEST WITH ZEBRAFISH

Measured conc. [mg/l]	% Mortality after:
	24h	48h	72h	96h
control	0	0	0	0
7.8	100	-	-	-
4.4	0	0	0	0
4*	0	0	0	0
2*	0	0	0	0
1*	0	0	0	0

* Nominal concentration, no supporting analysis was done. The concentrations were not used for calculation of the LC50 value.

Validity criteria fulfilled:

yes

Conclusions:

In a 96-h acute toxicity study according to OECD 203 with Danio rerio a 96-h LC50 of 5.9 mg/l was generated as the geometric mean of the highest concentration showing no lethal effect and the lowest concentration with 100% lethal effect. The test substance Aminoxide WS 35 is of moderate toxicity for freshwater fish.

Executive summary:

In a 96-h acute toxicity study according to OECD 203, Danio rerio were exposed to AMINOXIDE WS 35 at nominal concentrations of 0 (control), 1.0, 2.0, 4.0, 8.0, 10 mg test substance/l under semi-static conditions. The 96-h LC₅₀was generated as the geometric mean of the highest concentration showing no lethal effect and the lowest concentration with 100% lethal effect and determined as 5.9 mg a.i./l (measured conc.). There was high loss of test substance under test conditions (renewal after 48h) observed. Therefore the biological results were based on measured concentrations.
 
This study is classified as reliable with no restriction and satisfies the guideline requirements for an acute toxicity study with fish.

Results Synopsis
Test organism length: 1 – 3 cm
Test Type: Semi-static

96 h-LC₅₀: 5.9 mg/l (measured conc.)
Endpoint(s) Effected: mortality

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1999-12-06 - 1999-12-10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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: Aliquots of the stock solution were each separately dispersed in dechlorinated tap water and the volume adjusted to give the 0.18, 0.32, 0.56, 1.0, 1.8 and 3.2 mg a.i./L test concentrations

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: test material was prepared by direct solution in water. Test material was dissolved in dechlorinated tap water with the aid of ultrasonification for approx. 30 min. and the volume adjusted to 1 L

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Oncorhynchus mykiss
- Strain: Rainbow trout
- Source: Brow Well Fisheries Ltd. Hebden, Nr. Skipton UK
- Length at study initiation (length definition, mean, range and SD): mean standard length of 4.6 cm (sd=0.4)
- Weight at study initiation (mean and range, SD): mean zeight of 1.2 g (sd = 0.38)
- Feeding during test: no


ACCLIMATION
- Acclimation period: 14 November - 6 December 1999
- Acclimation conditions (same as test or not): the lightning cycle was controlled to give a 16 hrs light and 8 hrs darkness cycle with 20 minutes dawn and dusk transition periods
- Type and amount of food: commercial trout pellets

- Feeding frequency: 0.6 g body weight/liter
- Health during acclimation (any mortality observed):

QUARANTINE (wild caught)
- Duration: zero mortality in the 7 days prior to the start of the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

not stated

Test temperature:

14± 1°C

pH:

7.4-7.7

Dissolved oxygen:

9.2-9.8 mg O2/L

Salinity:

not stated

Nominal and measured concentrations:

range-finder: nominal concentrations of 1.0, 10 and 100 mg a.i./L
1st definitive study: 1.0, 1.8, 3.2, 5.6 and 10 mg a.i./L
2nd definitive study: 0.18, 0.32, 0.56, 1.0, 1.8, 3.2 mg a.i./L
Analysis of the test preparations at 0, 24 and 48h showed the measured concentrations to be within the range of 83% to 98% of nominal values with the exception of the 0.18, 0.32, 1.0 and 1.8 mg a.i./L test groups at 0hrs which showed measured values of 42%, 67%, 75% and 79% of nominal resp. These variable results were considered not to reflect the true dosed concentrations, but were the result of an unidentified technical error during the analytical procedure. Given that the 0 hr mean measured test concentrations and the 24h and 96h measured values were all within the range of 83% to 96% of nominal, the results are based on nominal concentrations only.

Details on test conditions:

TEST SYSTEM
- Test vessel: glass fiber tank with a single pass water renewal system
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 21 L
- Aeration: yes, via narrow bore glass tubes
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate): daily renewal
- No. of organisms per vessel:
- No. of vessels per concentration (replicates):
- No. of vessels per control (replicates):
- No. of vessels per vehicle control (replicates):
- Biomass loading rate:

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: the lighting cycle was controlled to give a 16 h light and 8 h darkness cycle with 20 min dawn and dusk transition periods
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Justification for using less concentrations than requested by guideline: /
- Range finding study
- Test concentrations: 1, 10, 100 mg a.i./L
- Results used to determine the conditions for the definitive study: yes

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.75 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.56-1.0

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.56 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: sun lethal effect of exposure was observed at the test concentration of 1.0 mg a.i./L. The response was fish swimming at the test concentration of 1.0 mg a.i./L

Reported statistics and error estimates:

Analysis of the mortality data was done by the trimmed Spearman-Karber method of Hamilton et al (1977) at 48h and the geometric mean method at 24, 72 and 96h based on nominal test concentrations

Validity criteria fulfilled:

yes

Conclusions:

The 96 h LC50 based on nominal test concentrations was 0.75 mg a.i./L with 95% confidence limits of 0.56-1.0 mg a.i./L.

Executive summary:

Analysis of the test preparations at 0, 24 and 96 hrs showed the measured test concentrations to be within the range of 83% to 93% of nominal values with the exception of the 0.18, 0.32, 1.0 and 1.8 mg a.i./L test groups at 0 hrs which showed measured values of 42%, 67%, 75% and 79% of nominal resp. These variable results were considered to reflect the true dosed test concentrations, but were the result of an unidentified technical error during the analytical procedure. Given the 0 hr mean measured concentratrions and the 24 hr and 96 hr measured values were all within the range of 83% to 93% of nominal, the results awere based on nominal test concentrations only.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-01-14 - 2013-01-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity 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 test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution was prepared by solving an appropriate amount of the test item in purified water and stirring carefully to avoid formation of air bubbles for about 2 hours at room temperature.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Oncorhynchus mykiss
- Strain: Rainbow trout
- Source: Sauerländer Forellen GmbH, Saalhauser Str. 8, 57368 Lennestadt-Gleierbrück, Germany
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD): The fish lengths were in the range of 5.0 – 5.7 cm.
- Weight at study initiation (mean and range, SD): The mean weight resulted in a loading of 0.6 g/L test medium

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

1 mmol/L

Test temperature:

14.8 °C until 15.2 °C

pH:

The pH in the test vessels was between 7.9 and 8.3

Dissolved oxygen:

oxygen saturation in all test vessels was between 94 % and 108 %

Salinity:

not stated

Nominal and measured concentrations:

A non-GLP pre-test was conducted using the following nominal concentrations: control, 1, 10, and 100 mg/L (corresponding to 0.36, 3.6, and 35.8 mg a.i./L). Based on the results of the range-finder test, the nominal concentrations of the test item used were 0.625, 1.25, 2.5, 5.0, and 10 mg/L (corresponding to 0.22, 0.45, 0.90, 1.79, and 3.58 mg a.i./L).

Details on test conditions:

TEST SYSTEM
- Test vessel: The test vessels were full glass aquaria of 28 L for keeping up to 20 litres of test solution. Glass vessels were pre-saturated with the test item. For this a volume of approximately 500 mL of test solution was filled into the glass vessel, was followed by gentle moving of the aquaria resulting in a complete coverage of the glass surface. Afterwards, the test media was discarded and the glass vessels were filled with the remaining solution.
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 28 L
- Aeration: The test vessels were slightly aerated via a glass capillary.
- Type of flow-through (e.g. peristaltic or proportional diluter):
- Renewal rate of test solution (frequency/flow rate): Test media were changed daily.
- No. of organisms per vessel: Seven fish each were exposed to five nominal concentrations for a period of 96 h and seven fish were used for the control

OTHER TEST CONDITIONS
- Photoperiod: The tanks were subjected to a light/dark cycle of 14/10 hours


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: Based on the results of the range-finder test, the nominal concentrations of the test item used were 0.625, 1.25, 2.5, 5.0, and 10 mg/L (corresponding to 0.22, 0.45, 0.90, 1.79, and 3.58 mg a.i./L).
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

1.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC10

Effect conc.:

1.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 1.5-2.2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 1.5-2.1

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.68 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0.54-0.79

Details on results:

- Other biological observations: Clinical signs were observed at 0.625, 2.5, 5.0 and 10 mg test item/L. Total fish mortality was observed at 5.0 and 10 mg test item/L.

Reported statistics and error estimates:

Where the test results show mortality around 50 % they will be statistically analysed to determine LC10 and LC50 values together with 95 % confidence intervals using Probit-analysis assuming log-normal distribution of the values.
If it is not possible to determine LC50 because no mortality was observed, the LC50 will be reported as being > highest concentration tested. The evaluation of the effects will be based on loading concentrations for the test solution preparations.
The computer program ToxRat will be used for statistical evaluations.

Validity criteria fulfilled:

yes

Conclusions:

The LC50 for acute toxicity of rainbow trout by amides, C12-C18 (even numbered), N[3-(dimethylamino)propyl], N’-oxides) is 1.8 mg test item/L, corresponding to 0.68 mg a.i./L. The NOEC is 1.3 mg test item/L (0.47 mg a.i./L).

Description of key information

One study (Teigeler, 2013), investigating the acute toxicity of the substance to fish according to OECD guideline 203, was considered as the key study for endpoint coverage. The 96h-LC50 based on measured geometric mean test concentrations was 0.68 a.i. mg/L with 95% confidence limits of 0.5-8 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 fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

0.68 mg/L

Additional information

Three studies provide information on the short-term toxicity of the substance to fish.

 

The study of Teigeler (2013) is selected as key study for endpoint coverage. The study was run according to OECD guideline 203 and EU guideline C.1. The 96h-LC50 based on measured geometric mean test concentrations was 0.68 mg a.i./L with 95% confidence limits of 0.5-8 mg a.i./L. The study is given a Klimisch score of 1 and was conducted under GLP.

 

Additionally, two other GLP studies were carried out according to OECD guideline 203 and are considered as supporting studies. The study of Kamp (1997) gave a 96h-LC50 of 2.06 mg a.i./L and was assigned a Klimisch score of 1, while the study of Wetton (2000) gave a 96h-LC50 of 0.75 mg a.i./L and was assigned a Klimisch score of 2.