Amines, C14-16 (even numbered)-alkyldimethyl, N-oxides

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

followed the principles of GLP, but some parts of the study were not fully conducted under GLP

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

not specified

Remarks:

but GLP conform

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: 126 deg C
- Boiling point: n/a
- Vapour pressure: 1.7E-06 and 7.5E-05 Pa
- Water solubility: 409 g/L
- Henry's law constant: 3.7E-09-1.2E-08 Pa m³/mol
- log Pow: <2.7
- pKa: 4.01
- Stability in water: stable
- Stability in light: stable
- pH dependance on stability: n/a

OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: readily biodegradable

Analytical monitoring:

not specified

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The substance was stirred with a glass stirrer and added to test water without any pretreatment.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Length at study initiation: 2.8 cm (2.5 - 3.1 cm)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

22 +/- 1 °C

pH:

7.6 - 8..1

Dissolved oxygen:

7.0 - 9.0 mg/L

Nominal and measured concentrations:

nominal concentrations: 0, 10, 100 mg/L

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: O2, pH and temperature were measured daily

EFFECT PARAMETERS MEASURED : mortality and behavior after 2, 4, 24, 48, 72 and 96 hours

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 - < 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Abnormal behavior in the concentration of 10 mg/L.

Conclusions:

In this study the test substance was found to be toxic to Danio rerio after 96 hrs at a nominal test iten concentration of 100 mg/L. The LC50 (96 hrs) is between 10 and 100 mg/L (equivalent to 3-30 mg AO/L). A NOEC could not be defined because all tested concentrations shows effects.

Executive summary:

The acute toxicity of the test substance to Danio rerio was dtermined in a 96 -hour static test according to the OECD Guideline 203.

The concentrations of the test item of 10 and 100 mg/L were tested in parallel with a control. There was no mortality at the control group. The tested concentration of 10 mg/L shows effects but no mortality. The highest tested concentration without lethality was 10 mg/L. The LC50 was between 10 and 100 mg/L (no calculation), equivalent to 3 -30 mg AO/L. The lowest tested concentration with 100 % effect (mortality) was 100 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no analysis of test concentrations during the study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

no

Analytical monitoring:

no

Details on sampling:

not applicable

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test solutions in both the preliminary and the definitive test were prepared from stock solutions of the test substance in deionised water. The test substance dissolved well and a clear and homogenous solution was obtained. The test concentrations were prepared by the addition of the respective amounts of stock solution to the Dutch Standard Water.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: no data
- Age at study initiation: no data
- Length at study initiation: approx. 2 cm
- Weight at study initiation: medium wetweight = 0.3 g/fish
- Feeding during test: no

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

not applicable

Test temperature:

21.3-21.6 deg C

pH:

7.5-8.2

Dissolved oxygen:

>7.3 mg/L

Salinity:

not measured

Nominal and measured concentrations:

Nominal concentrations:
Preliminary test: 0, 0.1, 1, 10, 32, 100 or 1000 mg/L (equivalent to 0, 0.0237, 0.237, 2.37, 7.58, 23.7 or 237 mg AO/L)
Definitive test: 0, 100, 180, 324, 583 or 1050 mg/L (equivalent to 0, 23.7, 42.66, 76.79, 138.17 or 248.85 mg AO/L)

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquarium
- Type: Covered with glass plate
- Material, size, headspace, fill volume: glass, 2 L capacity filled to 1 L (preliminary test) 5 L capacity filled to 3 L (definitive test)
- Aeration: yes
- Type of flow-through (e.g. peristaltic or proportional diluter): not applicable
- Renewal rate of test solution (frequency/flow rate): Renewed after 2 days (semi-static test)
- No. of organisms per vessel: 3 fish (preliminary test) 7 fish (definitive test)
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dutch Standard Water
- Total organic carbon: <2.0 mg/L
- Metals: < 10 ug Cu/L
- Conductivity: < 5 uS/cm

OTHER TEST CONDITIONS
- Adjustment of pH: not applicable
- Photoperiod: 12 hours ambient light per day provided by fluorescent tubes


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

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Justification for using less concentrations than requested by guideline: not applicable
- Range finding study
- Test concentrations: 0, 0.1, 1, 10, 32, 100 or 1000 mg/L (equivalent to 0, 0.0237, 0.237, 2.37, 7.58, 23.7 or 237 mg AO/L)
- Results used to determine the conditions for the definitive study: no mortality and no sublethal effects were observed at concentrations up to and including 100 mg/L (23.7 mg AO/L)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

134 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

31.8 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: at 180 mg/L the fish showed reduced activity before their death. At 100 mg/L no sublethal effects were observed.
- Mortality of control: There were no deaths among control animals in either the preliminary or definitive study

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

The LC50 was calculated with a programme (Akzo programme SKBT, version 1.0) using both the trimmed Spearman-Karber method (5.3) and the binomial test method (5.4).

Sublethal observations / clinical signs:

Table 1: Mortality in the definitive test

Mg test substance/L	No. of surviving animals
	Hours
	0	2	18	24	42	67	96
0	7	7	7	7	7	7	7
100	7	7	7	7	7	7	7
180	7	7	7*	4*	1*	0	0
324	7	5	0	0	0	0	0
583	7	0	0	0	0	0	0
1050	7	0	0	0	0	0	0

 

* Reduced activity

Validity criteria fulfilled:

yes

Executive summary:

The acute toxicity of C12 AO to Danio rerio was determined in a 96-hour semi-static test performed according to OECD TG 203. The fish (7 per group) were exposed at nominal concentrations of 0, 23.7, 42.66, 76.79, 138.17 or 248.85 mg AO/L. The 96-h LC50 was reported as 31.8 mg AO/L.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1987-06-22 to 1987-07-31

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

No test material analysis.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

No analysis of test substance

GLP compliance:

no

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: 126 deg C
- Boiling point: n/a
- Vapour pressure: 1.7E-06 and 7.5E-05 Pa
- Water solubility: 409 g/L
- Henry's law constant: 3.7E-09-1.2E-08 Pa m³/mol
- log Pow: <2.7
- pKa: 4.01
- Stability in water: stable
- Stability in light: stable
- pH dependance on stability: n/a

OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: readily biodegradable

Analytical monitoring:

no

Details on sampling:

No data

Vehicle:

no

Details on test solutions:

Loading of the test medium with the test sample was done 2 hours before the start of the test.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Forellenhof Fredesloh, Dr O Muller, 3413 Moringen 1
- Length at study initiation (length definition, mean, range and SD): 5 ± 1 cm

ACCLIMATION
- Acclimation period: 17 days
- Food: Trout food Pelletgr S 1 (Rheinkrone Kraft-futter)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No data

Hardness:

No data

Test temperature:

14.0 ± 0.5 degrees Centigrade at 0 hours
14.4 ± 0.9 degrees Centigrade at 24 hours
14.6 ± 0.3 degrees Centigrade at 48 hours
14.7 ± 0.3 degrees Centigrade at 72 hours
14.7 ± 0.4 degrees Centigrade at 96 hours

pH:

pH 8 throughout the period of the test

Dissolved oxygen:

Oxygen concentration was reported without units:
10.4 ± 0.3 at 0 hours
10.1 ± 0.4 at 24 hours
10.1 at 48 hours
10.1 ± 0.3 at 72 hours
10.1 ± 0.3 at 96 hours

Salinity:

No data

Nominal and measured concentrations:

Nominal concentrations of test substance: 0, 2.0, 3.0, 4.5, 6.7, 10.0, 15.0, 22.5, 33.5 and 50.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass tank
- Type (delete if not applicable): Closed via specially fitted glass plates
- Material, size, headspace, fill volume: Length 300 mm, width 200 mm, height 200 mm
- Aeration: Air was bubbled into the test medium at a rate of 1 to 3 bubbles per second through Pasteur pippettes directly connected with the aerator by flexible PVC tubes
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted water prepared by dechlorinating the city water supply via active charcoal filters served as the test medium

OTHER TEST CONDITIONS
- Feeding: Withdrawn 24 hours before the test started
- Photoperiod: 12 hours

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

42 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

12.6 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

A range finding test indicated a mortality of at least 50 mg/L. Regular inspections during the acute toxicity test gave the results shown in the table below.

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

The cumulative percentage mortality for the exposure period of 96 hours was plotted against the log of the concentration of the test substance. A line was fitted to the points and the concentration corresponding to 50 % mortality was read from that line.

Sublethal observations / clinical signs:

Cummulative mortality and (percentage mortality) at intervals during the test

Concentration [mg/L]	Time after test start [h]
	0	1	24	48	72	96
50.0	0	0	9	10	10	10
	(0)	(0)	(90)	(100)	(100)	(100)
33.5	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
22.5	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
15.0	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
10.0	0	0	0	0	0	6
	(0)	(0)	(0)	(0)	(0)	(60)
6.7	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
4.5	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
3.0	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
2.0	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)
Negative Control	0	0	0	0	0	0
	(0)	(0)	(0)	(0)	(0)	(0)

Validity criteria fulfilled:

yes

Remarks:

Control mortality < 10 %; dissolved oxygen concentration > 60 % of air saturation value; test material concentration not measured, but based on biodegradation studies expected to remain constant over the duration of the test

Conclusions:

The LC50 (96 h) was 42 mg/L test substance, equivalent to 12.6 mg AO/L.

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990-02-15 to 1990-03-09

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

no anlaysis of test substance concentration

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

not specified

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: Local aquarium retailer; Batch 20-2-90
- Length at study initiation: Mean length 2.5 to 3.2 cm
- Weight at study initiation: mean wet weight per fish was 2.8 g
- Feeding during test: None


QUARANTINE
- Duration: 12 days

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No data

Hardness:

No data

Test temperature:

During the test the temperature ranged from 21 - 23.4 ºC.

pH:

The variation during the test was in the range of 7.4 to 8.2.

Dissolved oxygen:

The oxygen concentration throughout the test ranged from 52 % of the air saturation value (at a concentration of 5.6 mg/L after 48 h) to 76 % of the air saturation value (at a concentration of 5.6 mg/L at 96 h). In general oxygen values were in the range of approximately 60 to 70 % of the air saturation value. Only in a few cases the oxygen concentration dopped below 60 % of the air saturation value. During the entire test period, no sign of oxygen shortage could be observed in the behaviour of the fish at any of the test concentrations.

Salinity:

No data

Nominal and measured concentrations:

Nominal:
Preliminary study: 1.0, 10 and 32 mg/L and 0.32, 1.0 and 3.2 mg/L.
Definitive test: 1.0, 1.8, 3.2, 5.6 and 10 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: 5 L glass aquaria
- Type: Closed; covered with a glass plate
- Fill volume: 3 L
- Aeration: The test medium was not aerated during the test
- Type of flow-through: Not applicable, test was semi-static
- Renewal rate of test solution: At 48 h
- No. of organisms per vessel: Preliminary test: 3; definitive test: 10.
- No. of vessels per concentration: No data
- No. of vessels per control (replicates): One
- Biomass loading rate: 1.0 g of biomass/L


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Synthetic water, Dutch Standard Water having a pH of approximately 8.2 and a hardness of 13 ºdH
- Total organic carbon: Not more than 2.0 mg/L
- Particulate matter: No data
- Metals: Not more than 10 μg copper
- Conductivity: Less than 5 μS/cm
- Culture medium different from test medium: No
- Intervals of water quality measurement: No data
Containing per litre of deionised water:
100 mg of NaHCO3
20 mg of KHCO3
200 mg of CaCl2.2H2O
180 mg of MgSO4.7H2O

OTHER TEST CONDITIONS
- Adjustment of pH: No data
- Photoperiod: 12 h ambient light per day, provided by fluorescent tubes.
- Light intensity: No data

EFFECT PARAMETERS MEASURED: pH and oxygen concentrations.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: Preliminary study: 1.0, 10 and 32 mg/L and 0.32, 1.0 and 3.2 mg/L; definitive test: 1.0, 1.8, 3.2, 5.6 and 10 mg/L.
- Results used to determine the conditions for the definitive study: 100 % mortality was observed after 24 h at 10 g/L and no mortality was observed at 1.0 mg/L after 96 h.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 3.5 - 7.1 mg/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.49 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

In the first preliminary test, 100 % mortality was observed after 24 h at 10 g/L and 32 mg/L and no mortality was observed at 1.0 mg/L after 96 h. A second preliminary test was then performed which showed no mortalities within 7 h at the concentrations up to and including 3.2 mg/L.
In the definitive test, the highest concentration of the test substance causing no morality after 96 h was 3.2 mg/L, whereas 10 mg/L produced 100 % mortality within 24 h. No other effects, such as deviations in the behaviour or the appearance of the fish were observed. The LC50 of the test substance (96 h) was calculated to be 5.0 mg/L with 95 % confidence limits of 3.5 and 7.1 mg/L. The 96 h LC50 of the active ingredient was calculated as 1.49 mg/L.

Results with reference substance (positive control):

The quality of the batch fish used in the test was further checked by means of a test with a reference substance (potassium dichromate). The results obtained are in accordance with the test results of an EEC study mentioning a concentration of 205.5 mg/L as LC50 48 h (mean value).

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

Mortality in the definitive test:

mg of test substance/L	Number of surviving animals
	0 h	24 h	48 h	72 h	96 h
Control	10	10	10	10	10
1.0	10	10	10	10	10
1.8	10	10	10	10	10
3.2	10	10	10	10	10
5.6	10	8	6	2	1
10	10	0

Reference substance mortality:

mg of test substance/L	Number of surviving animals
	0 h	24 h	48 h	72 h	96 h
Control	10	10	10	10	10
100	10	10	10	10	10
210	10	10	5	3	3
460	10	1	0

Quality criteria.

Mortality: The mortality of the control was less than 10 % and therefore this criterion was fulfilled.

Oxygen concentration: With the exception of the cases mentioned above, the quality criterion concerning the oxygen concentration > 60 % of the air saturation value throughout the test was fulfilled. Considering that no indication of oxygen shortage in the behaviour of the fish was observed, a harmful effect due to oxygen shortage can be excluded.

Biodegradability: It is likely that the concentration of the test substance remained constant during the test.

Validity criteria fulfilled:

yes

Conclusions:

Aromox DMB-W was tested for acute toxicity with Brachydanio rerio (zebra fish) under semi static conditions in accordance with EEC Method C1 and OECD test guideline 203. The LC50 (96 h) of the test substance was calculated to be 5.0 mg/L with 95 % confidence limits of 3.5 and 7.1 mg/L. The highest concentration of the test substance causing no mortality after 96 h amounted to 3.2 mg/L whereas 100 % mortality was produced within 24 h with 10 mg/L. No other effects such as deviations in the behaviour or appearance of the fish were observed. The 96 h LC50 of the active ingredient was calculated as 1.49 mg/L.

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no analysis of test concentrations during the study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Dilution in synthetic water
- Controls: Yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dutch Standard water (pH ca. 8.2, hardness of 11.7°dH) + 1L of deionised water.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None
- Stock solution: 1 g test substance/L deionised water

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Source: Not specified
- Age at study initiation (mean and range, SD): Not specified
- Length at study initiation (length definition, mean, range and SD): approx. 2 cm
- Weight at study initiation (mean and range, SD): approx 0.3 g (wet weight)
- Feeding during test: No

ACCLIMATION
- Acclimation period: Not specified

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Dutch Standard Water: 11.7°dH

Test temperature:

22.1 - 22.3°C

pH:

7.4 - 8.2

Dissolved oxygen:

≥ 8.4 mg/L (92% of air saturation value)

Nominal and measured concentrations:

Preliinary Test: 0.1, 1, 10, 32 and 100 mg/L (technical product)
Definitive Test: 1, 2, 4, 8, 16 mg/L (technical product)

Details on test conditions:

TEST SYSTEM
- Test vessel: Preliminary test: 2L aquaria, Definitive test: 5L aquaria
- Type (delete if not applicable): closed (glass plate)
- Fill volume: 3L test medium
- Aeration: Yes
- Renewal rate of test solution (frequency/flow rate): test fluids renewed after 2 days


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised water
- Total organic carbon: not more than 2.0 mg/L
- Metals: not more than 10 µg copper/L
- Conductivity: less than 5 µS/cm

OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 12 hours
- Light intensity: ambient (fluorescent tubes)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality at 24, 48, 72 and 96 hours

TEST CONCENTRATIONS
- Range finding study: yes
- Test concentrations: 0.1, 1, 10, 32 and 100 mg/L
- Results used to determine the conditions for the definitive study: Yes

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

10.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 8.5 - 12.3 mg/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.4 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Sublethal observations / clinical signs:

Mortality in preliminary test

mg of test substance*/L	Number of surviving animals
	Hours
	0	2	3	19	24	27	43	48	67	72	75	91	96
Control	3	3	3	3	3	3	3	3	3	3	3	3	3
0.1	3	3	3	3	3	3	3	3	3	3	3	3	3
1	3	3	3	3	3	3	3	3	3	3	3	3	3
10	3	3	3	3”	3’	2”	2”	2”	2”	2”	2”	2”	2”
32^+	3	1”	0

* = technical product

“ = reduced activity

‘ = one fish almost dead, reduced activity

+ = slight foaming

Mortality in definitive test

mg of test substance*/L	Number of surviving animals
	Hours
	0	3	19	24	26	42	48	50	67	70	73	90	96
Control	7	7	7	7	7	7	7	7	7	7	7	7	7
1	7	7	7	7	7	7	7	7	7	7	7	7	7
2	7	7	7	7	7	7	7	7	7	7	7	7	7
4	7	7	7	7	7	7	7	7	7	7	7	7	7
8	7	7	7	7	7	7	7	7	7	6	6	6	6
16^+	7	7’	0

* = technical product

‘ = reduced activity

+ = slight foaming

Validity criteria fulfilled:

yes

Conclusions:

The LC50 (96h) was calculated to be 10.3 mg/L equivalent to 2.4 mg AO/L.

Executive summary:

In a 96 h acute toxicity study Zebra fish (Brachydanio rerio) were exposed to C14 AO under semi-static conditions. The LC 50 (96h) value based on nominal concentration of the test material was 10.3 mg/L. The LC50 (96h) based on 100% active ingredient was 2.4 mg/L. This toxicity study is classified as acceptable and is similar to the OECD 203 Guideline.

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

no analysis of test substance concentration

Principles of method if other than guideline:

Groups of 10 fish were exposed to the test substance for 96 hours. Mortalities were recorded and the LC50 determined by Probit analysis.

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Leuciscus idus melanotus

Details on test organisms:

TEST ORGANISM
- Common name: Golden orfe
- Age at study initiation (mean and range, SD):
- Length at study initiation (length definition, mean, range and SD):
- Weight at study initiation (mean and range, SD): 1.3-7.1 g (mean 3.5 g, SD 1.4 g)

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20 +/- 0.5 deg C

pH:

Test solutions: 7.05-8.01
Controls solutions: 7.05 - 8.05

Dissolved oxygen:

6.2-8.65 mg/L

Nominal and measured concentrations:

0, 0.1, 0.8, 1, 2, 5, 10, 12.5, 31.5, 80, 100 mg/L (based on test substance)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

14.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.32 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Reference 7

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Length at study initiation: 2.80+/-0.16 cm

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96

Test temperature:

22 ± 1°C

pH:

7.8-8.1

Dissolved oxygen:

7.6-9.0 mg/L

Salinity:

no data

Nominal and measured concentrations:

0, 10, 100 mg/L nominal

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 - < 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 2.5 - < 25 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

All fish died within 2-4 h observation period at 100 mg/L test concentration

Executive summary:

In a study performed to OECD 203, the test material was dosed at 0, 10 or 100 mg/L (equivalent to 0, 2.5 or 25 mg AO/L) to Zebra fish for 96 h. No mortalities were observed at the 2.5 mg AO/L test concentration but all fish died within 2 - 4 hours of dosing with 25 mg AO/L. The LC50 is between 2.5 and 25 mg AO/L.

Reference 8

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1976-06-01

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

No measurement of test concentration

Qualifier:

according to guideline

Guideline:

other: APHA Standard Method (1970). Reliability, deviations and validity criteria evaluated against APHA Standard Method 8910 (1997).

Deviations:

yes

Remarks:

(10 fish per test concentration, not 20. Details on DO measurements not provided. No measurement of test concentration.)

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable
PHYSICO-CHEMICAL PROPERTIES
- Melting point: 126 deg C
- Boiling point: n/a
- Vapour pressure: 1.7E-06 and 7.5E-05 Pa
- Water solubility: 409 g/L
- Henry's law constant: 3.7E-09-1.2E-08 Pa m³/mol
- log Pow: <2.7
- pKa: 4.01
- Stability in water: stable
- Stability in light: stable
- pH dependance on stability: n/a

OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: readily biodegradable

Analytical monitoring:

no

Details on sampling:

Note: samples were collected, preserved, and sent to sponsor for analysis. However, details of the analytical measurement, and results, are not provided in this study report.
- Concentrations: Samples of 100 ml of test solutions were removed from selected concentrations (0.65, 2.8, 8.7mg/L) at various time intervals 0, 4, 8, 24, 48, 96h. in each test
- Sampling method: 100ml of samples were collected at each sampling period and added to 1ml of 37% formaldehyde
- Sample storage conditions before analysis: All water samples were shipped to the Procter & Gamble Company for chemical analyses

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:Reconstituted water used in the test system was prepared by adding 2mg potassium chloride, 30mg calcium sulfate, 30mg magnesium sulfate, and 48mg sodium bicarbonate per liter. The pH of the diluent was 7.1 and methyl orange alkalinity was 35ppm as CaCO3 and total hardness was 35mg/L
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: Control vessel contained 100 % reconstituted dilution water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill
- Strain: Not available
- Source: Commercial fish hatchery in Nebraska
- Age at study initiation (mean and range, SD): Not available
- Length at study initiation (length definition, mean, range and SD): mean length of 43mm
- Weight at study initiation (mean and range, SD): mean weight of 1.2g
- Method of breeding: Not available
- Feeding during test: Not available
- Food type: Not available
- Amount: Not available
- Frequency: Not available


ACCLIMATION
- Acclimation period: at least 24h in the water quality and test conditions
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: Not available
- Feeding frequency: Not available
- Health during acclimation (any mortality observed): Not available


QUARANTINE (wild caught)
- Duration: Not applicable
- Health/mortality: Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not applicable

Hardness:

35mg/L as CaCO3

Test temperature:

Static bioassays were conducted at 18 (+/-0.5) degree C in water bath

pH:

pH measured at 0, 48 and 96h pH ranged as follows:
Control: 7.1
For 0.65 mg/L :7.1
For 2.8 mg/L: 7-7.1
For 8.7 mg/L: 6.8-7.3

Dissolved oxygen:

DO levels ranged from initially 9.8 to 5.4 mg/L at the end of the test.

Salinity:

Not applicable

Nominal and measured concentrations:

Nominal laboratory exposure concentrations based on active ingredient were 0(control), 0.65, 1.2, 2.8, 4.9, 6.5, 8.7 mg/L.
No measurement of test concentrations

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed: Not available
- Material, size, headspace, fill volume: 5 gallon glass vessels
- Aeration: Not aerated
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Not applicable
- No. of organisms per vessel: Not available
- No. of vessels per concentration (replicates):Not available
- No. of vessels per control (replicates): Not available
- No. of vessels per vehicle control (replicates): Not available
- Biomass loading rate: Not applicable


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The reconstituted water used in the test was prepared by adding NaHCO3 (48 mg/L), CaSO4 (30 mg/L), MgSO4 (30 mg/L), and KCl (2mg/L) to deionized water.
- Total organic carbon: Not available
- Particulate matter: Not available
- Metals: Not available
- Pesticides: Not available
- Chlorine: Not available
- Alkalinity: 35ppm as CaCO3
- Ca/mg ratio: 30mg/L CaSO4 / 30mg/L MgSO4
- Conductivity: Not available
- Culture medium different from test medium: No
- Intervals of water quality measurement: Temperature and DO were measured initially and at the end of the test, pH was observed at 24 and 96 hours.


OTHER TEST CONDITIONS
- Adjustment of pH: Not applicable
- Photoperiod: Not available
- Light intensity: Not available


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality of Bluegill was measured at 24 and 96h


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.3 -2.3
- Justification for using less concentrations than requested by guideline: Not applicable
- Range finding study: No
- Test concentrations: Not applicable
- Results used to determine the conditions for the definitive study: Not applicable

Reference substance (positive control):

yes

Remarks:

p,p' DDT

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.13 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL 2.02-4.84mg/L. The study report included the endpoints as TL50 values. TL50 and LC 50 values are equivalent.

Details on results:

- Behavioural abnormalities: None
- Observations on body length and weight: None
- Other biological observations: Fish generally became dark and lethargic, lost equilibrium, and expired
- Mortality of control: None
- Other adverse effects control: None
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Mortality: Not available
- LC50: 0.007mg/L
- Other: 95%CL at 96h: 0.004 - 0.011mg/L

Reported statistics and error estimates:

LC50 values and it's 95% confidence limits for 96h were calculated by converting the test concentrations and corresponding mortalities to logs and probits, respectively. These values were utilized to calculated linear regression equation.

Validity criteria fulfilled:

yes

Remarks:

Criteria of Standard Method 8910: Mortality of controls <10%

Conclusions:

The 96 h LC50 of Alkyl dimethyl amine oxide to Bluegill was 3.13 mg a.i./L (based on nominal test concentration) with 95% CL of 2.02- 4.84mg/L.

Executive summary:

In a 96 hours acute toxicity study, Bluegill (Lepomis macrochirus) were exposed to Alkyl dimethyl amine oxide (NPS-72.004) at nominal concentrations of 0 (control), 0.65, 1.2, 2.8, 4.9, 6.5, 8.7 mg a.i./L under static conditions. No measurement of test concentration. The 96h LC50 value based on mortality was 3.13 mg a.i./L with 95%CL of 2.02 -4.84mg a.i. /L.

This toxicity study is classified as acceptable and satisfies the guideline requirement for the APHA Standard Method (1997) acute fish toxicity study.

Reference 9

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1975-01-10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

No measurement of test concentration.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: APHA Standard Method (1971). Reliability, deviations, and validity evaluated against APHA Standard Method 8910 (1997).

Deviations:

yes

Remarks:

(No measurements of test concentrations. Acclimation period was 24 h instead of at east 48 h. DO not measured.

Principles of method if other than guideline:

Not applicable

GLP compliance:

no

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
Not applicable
PHYSICO-CHEMICAL PROPERTIES
- Melting point: 126 deg C
- Boiling point: n/a
- Vapour pressure: 1.7E-06 and 7.5E-05 Pa
- Water solubility: 409 g/L
- Henry's law constant: 3.7E-09-1.2E-08 Pa m³/mol
- log Pow: <2.7
- pKa: 4.01
- Stability in water: stable
- Stability in light: stable
- pH dependance on stability: n/a

OTHER PROPERTIES (if relevant for this endpoint)
- Results of test for ready biodegradability: readily biodegradable

Analytical monitoring:

no

Details on sampling:

- Concentrations: Not available
- Sampling method: Not available
- Sample storage conditions before analysis: Not available

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Stock solutions of the toxicant were prepared by weighing out the compound in on an analytical balance and then bringing the stock solution to final volume with distilled water. Concentrations were based upon active ingradient.
- Eluate: Not applicable
- Differential loading: Not applicable
- Controls: Prepared using same dilution water
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): Not applicable
- Evidence of undissolved material (e.g. precipitate, surface film, etc): None

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Strain: Not available
- Source: National water quality laboratory, Duluth, Minnesota.
- Age at study initiation (mean and range, SD): 40-60 days old
- Length at study initiation (length definition, mean, range and SD): Maximun mean length: 33.6 mm
- Weight at study initiation (mean and range, SD): Mean weight: 0.31 g
- Method of breeding: Not available
- Feeding during test: Not available
- Food type: Not available
- Amount: Not available
- Frequency: Not available


ACCLIMATION
- Acclimation period: 24 hours
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: Not fed
- Feeding frequency: Not applicable
- Health during acclimation (any mortality observed): Not available


QUARANTINE (wild caught)
- Duration: Not applicable
- Health/mortality: Not applicable

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96

Remarks on exposure duration:

h

Post exposure observation period:

Not applicable

Hardness:

For Soft water: 48 mg/L as CaCO3
For Well water: 220 mg/L as CaCO3
For well water: 212 mg/L as CaCO3
For well water: 214 mg/L as CaCO3

Test temperature:

For all water types temperature was maintained at 22 degree C

pH:

For Soft water: 7.5
For Well water: 6.0
For well water: 7.8
For well water: 9.0

Dissolved oxygen:

Not available

Salinity:

Not applicable

Nominal and measured concentrations:

Test concentrations were not specified.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed: Not available
- Material, size, headspace, fill volume: Jars
- Aeration: Not available
- Type of flow-through (e.g. peristaltic or proportional diluter): Not applicable
- Renewal rate of test solution (frequency/flow rate): Not applicable
- No. of organisms per vessel: Not available
- No. of vessels per concentration (replicates): Not available
- No. of vessels per control (replicates): Not available
- No. of vessels per vehicle control (replicates): Not applicbale
- Biomass loading rate: Not available


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: 4 different dilution waters were used. One was a reconstituted water; the other 3 were partially softened well water, buffered to different pH's. See Table below for details of these 4 waters.
- Total organic carbon: Not available
- Particulate matter: Not available
- Metals: Not available
- Pesticides: Not available
- Chlorine: Not available
- Alkalinity: See Table below
- Ca/mg ratio: Not available
- Conductivity: Not available
- Culture medium different from test medium: Not available
- Intervals of water quality measurement:


OTHER TEST CONDITIONS
- Adjustment of pH: Not applicable
- Photoperiod: Not available
- Light intensity: Not available


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality was observed at 24, 48 and 96 hours


TEST CONCENTRATIONS
- Spacing factor for test concentrations: Not available
- Justification for using less concentrations than requested by guideline: Not available
- Range finding study: Yes
- Test concentrations: Not available
- Results used to determine the conditions for the definitive study: Not available

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.46 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 3.29-3.63 mg/L; Reconstituted water (pH 7.5)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.18 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 2.83-3.58 mg/L; Well water (pH 6.0)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.67 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 2.50-2.87 mg/L; Well water (pH 7.9)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.41 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 3.22-3.60 mg/L; Well water (pH 9.0)

Details on results:

- Behavioural abnormalities: None
- Observations on body length and weight: None
- Other biological observations: None
- Mortality of control: Not available
- Other adverse effects control: Not available
- Abnormal responses: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None
- Effect concentrations exceeding solubility of substance in test medium: None

Results with reference substance (positive control):

Not available

Reported statistics and error estimates:

LC50 and its 95% CL were determined by moving average (Thompson, 1947)

Validity criteria fulfilled:

yes

Remarks:

Criteria of Standard Method 8910: Mortality of controls <10%

Conclusions:

The nominal 96 h acute toxicity of Alkyl Dimethyl Amine Oxide to fathead minnow ranged from 2.67 to 3.46 mg a.i./L in 4 different dilution waters.

Executive summary:

In a 96 h acute toxicity study, Fathead minnow (Pimephales promelas) were exposed to Alkyl Dimethyl Amine Oxide under static conditions. The 96 h LC50 value based on nominal concentration of active ingradient ranged from 2.67 to 3.46 mg a.i./L as follows:

Reconstituted Water, pH 7.5       3.46 mg/L

Well water, pH 6.0                     3.18 mg/L

Well water, pH 7.9                     2.67 mg/L

Well water, pH 9.0                     3.41 mg/L

This toxicity study is classified as acceptable and satisfies the guideline requirements for the APHA Standard Method 8921 (1997) Fish acute toxicity study.

Reference 10

Endpoint:

short-term toxicity to fish

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

REPORTING FORMAT FOR THE CATEGORY APPROACH

1. HYPOTHESIS FOR THE CATEGORY APPROACH (ENDPOINT LEVEL)
Please refer to the Amine Oxide Category justification attached in Section 13

2. CATEGORY APPROACH JUSTIFICATION (ENDPOINT LEVEL )
Study summaries of the available data for category members are provided in IUCLID Section 6.1.1. These data demonstrate an increase in acute toxicity to fish as chain length increases moving from C12 AO to C14 AO and C12-14 AO. The acute toxicity to fish of C12-18 AO is of the same order of magnitude as C12-14 AO.

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes

Analytical monitoring:

not specified

Details on sampling:

N/A

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The sample was described as soluble in water. After addition to water the sample dispersed well throughout the test vessel. Dilution media was freshwater from the laboratory water supply.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Brow Well Fisheries Ltd, Hebden, Skipton, North Yorkshire.
- Age at study initiation (mean and range, SD): Juvenile


ACCLIMATION
- Acclimation period: Duration not specified
- Acclimation conditions: Stock juvenile rainbow trout were held in freshwater maintained at 15 ± 2 ºC by a chiller unit and aerated via air diffusers attached to the laboratory air supply system. Feeding was stopped when juvenile rainbow trout were transferred to test tanks 24 h before the start of the test.
- Type and amount of food: No data
- Health during acclimation: Free of malformations and in good health.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

No data

Hardness:

No data

Test temperature:

Nominal temperature of 15 ± 2.0 ºC.
Measured temperature of 15.0 to 15.5 ºC.

pH:

Recorded at 24 h intervals for control and test media:
Control - Old media: 8.19 to 8.27.
Control - New media: 8.32.
5.62 mg/L - Old media: 8.26
10mg/L - New media: 8.41
10mg/L - Old media: 8.28 to 8.36

Dissolved oxygen:

Recorded at 24 h intervals for control and test media: 100 %

Salinity:

No data

Nominal and measured concentrations:

Nominal concentrations
Range finder: 0, 10 and 100 mg/L.
Definitive study: 1.0, 1.78, 3.16, 5.62 and 10 mg/L.

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: 10 L/concentration
- Renewal rate of test solution: Replaced at 48 h
- No. of organisms per vessel: Seven
- No. of vessels per concentration: Single


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 10 and 100 mg/L
- Results used to determine the conditions for the definitive study: Due to unexplained control mortality in the rangefinder study the data was deemed unusable. From the results observed in a Daphnia magna study, a definitive range of 10 to 100 mg/L was set, however this test range produced 100 % mortality. The test was then repeated at a range of 1 to 10 mg/L.

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.26 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

All animals in the control vessels survived the 96 h test period. After 96 h exposure to the test material, the LC50 was calculated to be 4.2 mg/L . The LC50 after 72 h was calculated to be 4.6 mg/L. The 24 and 48 h LC50 values were both calculated to be 7.5 mg/L in this study.
There was some evidence of a dose response.

Results with reference substance (positive control):

No data

Reported statistics and error estimates:

No data

Sublethal observations / clinical signs:

Mortality data for juvenile rainbow trout exposed to control and test media containing Mackamine CS lot HG-161 (cocamine oxide) in a definitive study:

Test material concentration (mg/L)		Exposure period (h)
		0	24	48	72	96
0	Alive	7	7	7	7	7
	Dead	0	0	0	0	0
1.00	Alive	7	7	7	7	7
	Dead	0	0	0	0	0
1.78	Alive	7	7	7	7	7
	Dead	0	0	0	0	0
3.16	Alive	7	7	7	7	7
	Dead	0	0	0	0	0
5.62	Alive	7	7	7	6	0
	Dead	0	0	0	1	7
10.0	Alive	7	0	0	0	0
	Dead	0	7	7	7	7

Conclusions:

A sample of Mackamine CS lot HG-161 (cocamine oxide) was assessed for its potential toxicity to juvenile rainbow trout in a semi-static test over 96 h in accordance with OECD guideline 203. The highest concentration producing no mortalities was 3.16 mg/L; the lowest concentration producing 100 % mortality was 5.62 mg/L. The 96 h LC50 was 4.2. There was some evidence of a dose response.

Description of key information

No acute fish toxicity studies have been performed using C14 -16 AO. Data are available for category members C12 AO, C14 AO, C12 -14 AO and C12 -18 AO. The LC50 (96 h) for fish is 1.26 mg AO/L, based on the lowest LC50 derived from the key study performed using C12-18 AO.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.26 mg/L

Additional information

No acute fish toxicity studies have been performed using C14 -16 AO. Data are available for category members C12 AO, C14 AO, C12-14 and C12-18 AO. These data demonstrate an increase in acute toxicity to fish as chain length increases moving from C12 AO to C14 AO and C12-14 AO. The acute toxicity to fish of C12-18 AO is of the same order of magnitude as C12-14 AO.

Three reliable studies are available for C12-18 AO. In the key study, performed according to OECD TG 203 under GLP [Rowlands (2005)] Rainbow trout (Oncorhynchus mykiss) were exposed to C12-18 AO under semi static conditions for 96 hours. The LC50 (96 h) was 1.26 mg AO/L, based on nominal concentrations. In a supporting study [Mark & Meuwsen (1990)] performed according to OECD TG 203 under GLP, Danio rerio were exposed to C12-18 AO under semi static conditions for 96 hours. The LC50 (96 h) based on nominal concentrations was 1.49 mg AO/L. In the third study [Hoechst (1977)] Golden orfe were exposed to C12-18 AO for 96 hours. The LC50 (96 h) was 4.32 mg AO/L based on nominal test concentrations.

There are four reliable studies available for C12-14 AO. Iwan GR et al (1975) exposed Fathead minnow (Pimephales promelas) to C12-14 AO under static conditions for 96 hours. The LC50 (96 h) based on nominal concentrations of amine oxide ranged from 2.67 to 3.46 mg AO/L depending on the source and pH of the water used. In a supporting study Bluegill (Lepomis macrochirus) were exposed to C12-14 AO under static conditions for 96 hours. The LC50 value was reported as 3.13 mg AO/L [Macek & Sleight (1972)]. Exposure of rainbow trout (Oncorhynchus mykiss) to C12-14 AO under static conditions for 96 hours resulted in a LC50 of 12.6 mg AO/L [Dommrose AM (1987)], whilst exposure of zebra fish (Danio rerio) to C12-14 AO under static conditions resulted in a 96 -h LC50 of 3 -30 mg AO/L [Hoechst AG (1987)].

In addition, supporting studies are available for the following category members:

C12 AO: 96-h LC50 = 31.8 mg AO/L (Danio rerio) [Mark & Arends (1992)]

C14 AO: 96-h LC50 = 2.4 mg AO/L (Danio rerio; OECD TG 203) [Mark (1992)]; 96-h LC50 = > 2.5 - < 25 mg AO/L (Danio rerio; OECD TG 203) [Jung (1987)]