(2-hydroxy-3-sulphopropyl)dimethyl[3-[(1-oxododecyl)amino]propyl]ammonium hydroxide

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:

key study

Study period:

22 november 2012 - 13 march 2013

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

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Duplicate samples were taken from each treatment just before the start of the first and of the last renewal periods (Days 0 and 3). At the end of these renewal periods (Days 1 and 4), additional samples were taken for determination of the stability of the test item during the renewal periods of 24 hours. All samples were taken from the approximate center of each aquarium without mixing the test medium, and were stored deep-frozen (at about -20 °C) immediately after sampling. The concentrations of the test item were determined in one of the duplicate test medium samples from the nominal test concentrations of 28.1 mg/L from all sampling times.
From the control samples, only one of the duplicate samples of the freshly prepared test medium was analyzed per sampling time. The control samples of the aged test medium were not analyzed.

Vehicle:

no

Details on test solutions:

At the start of the test and before each test medium renewal, the test medium of nominal 28.1 mg/L was freshly prepared by completely dissolving 0.1407, 0.1407, 0.1404 and 0.1405 mg of the test item in five liter of test water under intense stirring for 15 minutes at room temperature, respectively. The test medium was freshly prepared just before introduction of the fish (= start of the exposure and prior to each test medium renewal).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish.
- Strain: Danio rerio.
- Source: breeding culture at Harlan Laboratories.
- Length at study initiation (length definition, mean, range and SD): 2.81 ± 0.14 cm (Mean ± SD).
- Weight at study initiation (mean and range, SD): 0.15 ± 0.03 g (Mean ± SD).
- Feeding during test: no.

ACCLIMATION
- Acclimation period: one week prior to test.
- Acclimation conditions: same as test.
- Type and amount of food: commercial fish diet.
- Health during acclimation: during holding and acclimatization no mortality observed and all fish were healthy.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

125 mg/L as CaCO3

Test temperature:

21°C

pH:

7.1 to 7.5

Dissolved oxygen:

At least 8.1 mg/L (corresponding to at least 90% oxygen saturation)

Nominal and measured concentrations:

Nominal concentration: 28.1 mg test item/L (limit test) corresponding to 12.9 mg solid content/L and 10 mg active content/L

Details on test conditions:

TEST SYSTEM
- Test vessel: one glass vessel with 5 liters of test medium for each treatment.
- Type: open.
- Aeration: slight aeration during the test.
- Renewal rate of test solution (frequency/flow rate): test media was renewed daily during the test period of 96 hours.
- No. of organisms per vessel: 7.
- No. of vessels per concentration (replicates): 1.
- No. of vessels per control (replicates): 1.
- Biomass loading rate: 0.21 g fish wet weight per liter test medium.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted test water. It consisted of analytical grade salts dissolved in purified water.
- Alkalinity: 0.4 mmol/L.
- Ca/Mg ratio: 4:1.
- Na/K ratio: 10:1.
- Intervals of water quality measurement: the water temperature, pH values and oxygen concentrations were measured for each treatment at the start of the test and once daily during the test in the freshly prepared and old test media.

OTHER TEST CONDITIONS
- Adjustment of pH: no.
- Photoperiod: 16 hour light to 8-hour dark photoperiod, with a 30-minute transition period was used.
- Light intensity: 140 to 480 Lux.

EFFECT PARAMETERS MEASURED: the test fishes were observed for mortality and visible abnormalities after approximately 4, 24, 48, 72 and 96 hours test duration.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: limit test / one concentration tested.
- Range finding study: no.
- Test concentrations: 28.1 mg/L (test item).

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 28.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 12.9 mg/L

Nominal / measured:

nominal

Conc. based on:

other: solid content

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 28.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 12.9 mg/L

Nominal / measured:

nominal

Conc. based on:

other: solid content

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Details on results:

Behavioural abnormalities: no.
Mortality of control: no.

In the test medium sample taken at the start and end of the first and last test medium renewal periods, the measured test item concentration ranged between 94 and 118% of the nominal value (less than 20% of deviation from nominal concentration). The oxygen concentration was at least 8.1 mg/L (see table 3) corresponding to at least 90% oxygen saturation.

All the validity criteria were fulfilled:
- mortality of controls is less than 10% at the end of the test;
- test conditions (pH, temperature) were maintained during the test (see tables 1 and 3);
- the test item concentration was maintained during the test (less than 20% of deviation from nominal concentration);
- the oxygen saturation was maintained during the test and was above 60% of oxygen saturation.

Sublethal observations / clinical signs:

Table 1     pH in the Freshly Prepared and Old Treatment Media

Nominal test item concentration (mg/L)	Exposure time
	0 h	24 h		48 h		72 h		96 h
	new	old	new	old	new	old	new	old
Control	7.3	7.2	7.2	7.5	7.4	7.4	7.3	7.2
28.1	7.3	7.2	7.1	7.5	7.4	7.4	7.3	7.2

  

Table 2     Oxygen Concentrations (mg/L) in the Freshly Prepared and Old Treatment Media

Nominal test item concentration (mg/L)	Exposure time
	0 h	24 h		48 h		72 h		96 h
	new	old	new	old	new	old	new	old
Control	9.8	8.4	9.6	8.5	9.9	8.4	9.7	8.2
28.1	9.7	8.4	9.7	8.5	9.9	8.5	9.6	8.1

  

Table 3     Temperatures (°C) in the Freshly Prepared and Old Treatment Media

Nominal test item concentration (mg/L)	Exposure time
	0 h	24 h		48 h		72 h		96 h
	new	old	new	old	new	old	new	old
Control	21	21	21	21	21	21	21	21
28.1	21	21	21	21	21	21	21	21

  

Validity criteria fulfilled:

yes

Conclusions:

The test item exhibited no adverse effects on zebra fish at the nominal concentration of 28.1 mg/L (corresponding to 12.9 mg solid content/L and 10 mg active content/L).

Executive summary:

The short-term aquatic toxicity of the REACh substance (2-Hydroxy-3-sulphopropyl) dimethyl[3-[(1-oxododecyl) amino]propyl]ammonium hydroxide (EC 242-893-1) was investigated in a GLP-compliant study performed with the freshwater fish species Danio rerio (zebra fish) in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test) and EU Method C.1. The test item exhibited no adverse effects on zebra fish at the nominal concentration of 28.1 mg/L (limit test) and the 96h-LC50 value is reported to be greater than 28.1 mg/L (corresponding to 12.9 mg solid content/L and 10 mg active content/L).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

02 june 2009 - 13 july 2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Despite the lack of analytical monitoring data, it can be reasonably assumed that the test concentrations remained stable over the test duration from the analytical measurements reported in the long-term toxicity test to Daphnia magna performed on a structural analogue substance (Höger, 2013).

Qualifier:

according to guideline

Guideline:

other: OSPAR / PARCOM Protocols on methods for the testing of chemicals used in the offshore Industry 2006

Deviations:

yes

Remarks:

fish length and loading rate above recommended maximum values

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Concentrations: 0 (control) and 10 mg test item/L.
- Method: samples were taken after 24, 48, 72 and 96 hours in order to observe and record mortalities. Water quality determinations for each test and control solution were carried out immediately prior to initiating the test, after 24 hours, before and after the 48 hour change of test solutions, after 72 hours and at the end of the 96 hour exposure period.
- Reference vessels: none.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: a stock solution of 1000 mg/L (nominal) was prepared with dilution water, shaken vigorously and allowed to stand for four hours. For each nominal concentration the required amount of homogenized stock solution was added to 10 litres of dilution water. Dilution water is a dechlorinated water with Tropic Marin artificial sea salt formulated by Tropical Marine Centre Limited added to a final salinity of 33 to 35 g/L sodium chloride. Test solutions were replaced after 48 hours up to the maximum exposure period of 96 hours.
- Controls: culture medium without test item.
- Evidence of undissolved material (e.g; precipate, surface film, etc): no.

Test organisms (species):

other: Scophthalmus maximus (turbot)

Details on test organisms:

TEST ORGANISM
- Common name: turbot.
- Strain: Scophthalmus maximus.
- Source: France Turbot, Noirmoutiers, France.
- Loading rate: 2.08 g fish/litre.
- Feeding during test: no.
- Length at study initiation: mean length of 58.7 mm.
- Weight at study initiation: mean weight of 2.97 g.

ACCLIMATION
- Acclimation period: ca. 2 month.
- Acclimation conditions: Scophthalmus maximus were maintained in dechlorinated mains water with Tropic Marin artificial sea salt formulated by Tropical Marine Centre Limited at 14.5 to 15.5°C, above 98% dissolved oxygen and under a photoperiod of 16 hours light and 8 hours dark.
- Type and amount of food: fed with Le Gouessant Turbot Label Rouge at a rate of approximately 2% body weight per day until 24 hours before test initiation.
- Feeding frequency: daily.
- Health during acclimation (any mortality observed): 0.4% mortality during acclimatation period.

Test type:

semi-static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

no data

Test temperature:

15.0 - 15.5°C

pH:

8.2 - 8.3

Dissolved oxygen:

83 - 101%

Salinity:

32 ‰

Nominal and measured concentrations:

Nominal concentration: 10 mg test item/L (corresponding to 4.8 mg solid content/L).

Details on test conditions:

TEST SYSTEM
- Test vessel: 12 litre volume aquaria
- Type: open
- Aeration: no data
- Renewal rate of test solution (frequency/flow rate): test solutions replaced after 48 hours up to the maximum exposure period of 96 hours.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 2.08 g fish per litre

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water is a dechlorinated water with Tropic Marin artificial sea salt formulated by Tropical Marine Centre Limited added to a final salinity of 33 to 35 g/L sodium chloride.
- Culture medium different from test medium: No
- Intervals of water quality measurement: each 24 hours up to the end of the test after 96 hour exposure period.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours light and 8 hours dark

EFFECT PARAMETERS MEASURED: The fish were observed for mortality and abnormal behaviour after 24, 48, 72 and 96 hours exposure periods.

TEST CONCENTRATIONS
Limit test at 10 mg/L (nominal)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 4.8 mg/L

Nominal / measured:

nominal

Conc. based on:

other: solid content

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Details on results:

Behavioural abnormalities: no.
Mortality of control: no.

The following validity criteria were fulfilled:
- mortality of controls is less than 10% at the end of the test;
- test conditions (pH, temperature, salinity) were maintained during the test;
- the oxygen saturation was maintained during the test and was above 60% of oxygen saturation.

There is a lack of information of data relating to the analytical monitoring showing that the concentrations of the test item were maintained during the test. However, according to the long-term toxicity test to Daphnia performed on a very similar substance, the substance is stable during 48 hours. Thus, we can consider that in a short-term study, the substance is also stable.

The largest fish tested was 65 mm and the loading rate was 2.08 g fish per litre while the guideline recommends a maximum fish length of 60 mm and a loading rate of 1 g fish per litre of test solution. Nevertheless, water qualities remains within recommended limits and no mortalities or abnormal behavior of the Turbot was recorded. Thus, this deviation is considered not to have adversely affected the data.

For details see tables 1 to 4 in ‘Any other information on results incl. tables’.

Sublethal observations / clinical signs:

Table 1: Summary of pH values

Exposure period (hours)	Concentration (mg/L)
	Control	10.0
0	8.2	8.2
24	8.2	8.2
48	8.2	8.2
	8.3	8.2
72	8.3	8.3
96	8.2	8.3

 

Table 2: Summary of dissolved oxygen values (% ASV)

Exposure period (hours)	Concentration (mg/L)
	Control	10.0
0	101	100
24	100	99
48	99	98
	101	100
72	99	99
96	83	92

 

Table 3: Summary of temperature values (°C)

Exposure period (hours)	Concentration (mg/L)
	Control	10.0
0	15.0	15.0
24	15.5	15.5
48	15.0	15.0
	15.0	15.0
72	15.5	15.5
96	15.5	15.5

 

Table 4: Summary of salinity (‰)

Exposure period (hours)	Concentration (mg/L)
	Control	10.0
0	32	32
24	32	32
48	32	32
	32	32
72	32	32
96	32	32

 

Validity criteria fulfilled:

yes

Conclusions:

The test item exhibited no adverse effects on turbot at the nominal concentration of 10 mg/L (corresponding to 4.8 mg solid content/L).

Executive summary:

The short-term aquatic toxicity of the REACh substance (2-Hydroxy-3-sulphopropyl) dimethyl[3-[(1-oxododecyl) amino]propyl]ammonium hydroxide (EC 242-893-1) was investigated in a GLP-compliant study performed with the marine fish species Scophthalmus maximus (turbot) in accordance with OSPARCOM guidelines. The test item exhibited no adverse effects on turbot at the nominal concentration of 10 mg/L (limit test) and the 96h-LC50 value is reported to be greater than 10 mg/L (corresponding to 4.8 mg solid content/L).

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19 january 2009 - 24 march 2009

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Despite the lack of analytical monitoring data, it can be reasonably assumed that the test concentrations remained stable over the test duration from the analytical measurements reported in the long-term toxicity test to Daphnia magna performed on a structural analogue substance (Höger, 2013).

Qualifier:

according to guideline

Guideline:

other: OSPARCOM (1995) (amended March 2005) - Protocol for a fish acute toxicity test

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: the test solution was prepared without any organic solvent.
- Control: one replicate.
- Reference vessels: no reference vessels.
- Evidence of undissolved material (e.g. precipitate, surface film, etc): none.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: the test item is soluble in water and was added to the test system directly via seawater.
- Stock solution: no. At the beginning of the test, 0.03884 g of test item was diluted to 23000 ml of treated seawater. The same tested solution was generated at 48h.
- Controls: no data.
- Evidence of undissolved material (e.g; precipate, surface film, etc): no.

Test organisms (species):

other: Scophthalmus maximus (turbot)

Details on test organisms:

- Common name: turbot.
- Strain: Scophthalmus maximus.
- Source: Ardtoe Hatchery.
- Loading density (fish/litre): 0.878.
- Feeding during test: no.

ACCLIMATION
- Acclimation period: 24 days.
- Acclimation conditions: Scophthalmus maximus were held under flow-through conditions in 200 litre capacity polythene tanks, containing 1 µm filtered ultra-violet treated seawater and supplied with gentle aeration.
- Type and amount of food: fed food in pellet form ad libitum.
- Feeding frequency: twice daily.
- Health during acclimation (any mortality observed): 0% mortality.

Test type:

semi-static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

no data

Test temperature:

14.3-14.8°C

pH:

7.65-7.99

Dissolved oxygen:

89-97%

Salinity:

37-38 ‰

Nominal and measured concentrations:

nominal concentration: 1.69 mg/l

Details on test conditions:

TEST SYSTEM
Test vessel:
- Material, size, headspace, fill volume: 30 litre capacity moulded soda-lime glass tanks.
- Aeration: gentle aeration.
- Renewal rate of test solution: at 48h.
- No. of organisms per vessel: 7.
- No. of vessels per concentration (replicates): 1.
- No. of vessels per control (replicates): 1.
- Biomass loading rate: 0.878 g/L.

TEST MEDIUM / WATER PARAMETERS
- Source of dilution water: natural local seawater pumped to holding tanks from Scapa Flow, Orkney.
- Treatment process: sand filtered to remove particulate, filtered to 1 µm, heated to 16°C, UV treated, degassed and held at a quantity of 3000 to 5000 litres.
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no.
- Photoperiod: constant illumination.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1.69 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.51 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks:

no mortality observed at the limit test concentration

Details on results:

- Behavioural abnormalities: no
- Observations on body length and weight: measured at the end of the test (see table 2 above)
- Other biological observations: no
- Mortality of control: no
- Other adverse effects control: no
- Abnormal responses: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: -

- Total alive fish afetr 96h exposure to teste material: 7/7

- The following validity criteria are fulfilled:
* Mortality of controls is less than 10% at the end of the test
* Test conditions (pH, temperature, salinity) were maintained during the test
* The oxygen saturation was maintained during the test and was above 60% of oxygen saturation
* There is a lack of information of data relating to the analytical monitoring showing that the concentrations of the test item were maintained during the test. However, the test has been performed under semi-static conditions and according to the long-term toxicity test to Daphnia on a very similar substance, the substance is stable during 48 hours. Thus, we can consider that in a short-term study, the substance is also stable.
For details see table 1 in ‘Any other information on results incl. tables’

Sublethal observations / clinical signs:

Table 1: Summary of guideline criteria and values during test

 

Parameter	Guideline criterion	Observed values
Control pH over 96h	6-9	7.66 – 8.00
pH over 96h	6-9	7.65 – 7.99
Temperature (°C)	15 ±1.5	14.3 – 14.8
Dissolved oxygen (%)	≥60	89 – 97
Salinity (‰)	36±4	37 – 38
Control mortality (%)	≤10	0
Maximum loading of fish/litre	1g	0.878g

 

Validity criteria fulfilled:

yes

Conclusions:

The test item exhibited no adverse effects on turbot at the nominal concentration of 1.69 mg/L (corresponding to 0.51 mg active content/L).

Executive summary:

The short-term aquatic toxicity of the REACh substance (2-Hydroxy-3-sulphopropyl) dimethyl[3-[(1-oxododecyl) amino]propyl]ammonium hydroxide (EC 242-893-1) was investigated in a GLP-compliant study performed with the marine fish species Scophthalmus maximus (turbot) in accordance with OSPARCOM guidelines. The test item exhibited no adverse effects on turbot at the nominal concentration of 1.69 mg/L (limit test) and the 96h-LC50 value is reported to be greater than 1.69 mg/L (corresponding to 0.51 mg active content/L).

Description of key information

The short-term aquatic toxicity of the REACh substance (2-Hydroxy-3-sulphopropyl) dimethyl[3-[(1-oxododecyl) amino]propyl]ammonium hydroxide (EC 242-893-1) was investigated in a GLP-compliant study performed with the freshwater fish species Danio rerio (zebra fish) in accordance with OECD Guideline 203 (Fish, Acute Toxicity Test) and EU Method C.1 (Höger, 2013). The test item exhibited no acute toxic effects on zebra fish at the nominal concentration of 28.1 mg/L (limit test) and the 96h-LC50 value is reported to be greater than 28.1 mg/L (corresponding to 12.9 mg solid content/L and 10 mg active content/L).

The short-term aquatic toxicity of the REACh subsance was otherwise investigated in two GLP-compliant studies performed with the marine fish species Scophthalmus maximus (turbot) in accordance with OSPARCOM guidelines (Richardson, 2009; Sacker, 2009). The 96h-LC50 is reported to be greater than 1.69 mg/L (corresponding to 0.51 mg active content/L) and 10 mg/L (corresponding to 4.8 mg solid content/L), respectively.

NB. In case the solid content of the test item was not reported in a study report, ECx values were expressed in active content. If relevant, these values were used for the chemical safety assessment (worst-case values).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

12.9 mg/L

Marine water fish

Marine water fish

Effect concentration:

4.8 mg/L

Additional information

The short-term toxicity to fish of the substance EC 242-893-1 was investigated in three GLP-compliant studies (Höger, 2013; Richardson, 2009; Sacker, 2009) performed in accordance with standard methods, with no or minor deviations. These studies are considered as reliable (Klimisch 1 - with analytical monitoring - or Klimisch 2 - without analytical monitoring) and were selected as key studies for the endpoint.