Reaction mass of morpholine and 6-[(p-tosyl)amino]hexanoic acid, compound with morpholine (1:1)

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:

From January 18, 2007 to December 06, 2007

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:

other: PARCOM Marine Fish, Acute Toxicity Method

Version / remarks:

1995

Deviations:

no

Principles of method if other than guideline:

Test species Scophthalmus maximus is a standard species for OSPAR HMCS since it is a common species in the North Atlantic, Baltic Sea and Mediterranean Sea.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: p-Toluene sulphonamido caproic acid morpholine salt at a nominal concentration of 7.51 mg/L was diluted in artificial seawater (Prodac Ocean Fish Marine Salt), in which it was soluble and reportedly dispersed well. Each vessel contained a 10 litre volume of test or control media
- Controls: Artificial seawater

Test organisms (species):

other: Scophthalmus maximus

Details on test organisms:

TEST ORGANISM
- Common name: Juvenile turbot (Scophthalmus maximus)
- Source: Manx Mariculture Ltd, Castletown, Isle of Man

ACCLIMATION
- Acclimation conditions: Animals were maintained as detailed in STL Runcorn SOP I.16, with stock juvenile turbot held in artificial seawater (STL Runcorn SOP III.12) kept at 15 +/- 2 °C by a chiller unit and aerated via air diffusers attached to the laboratory air supply system
- Type and amount of food during acclimation: Animals were fed as detailed in STL Runcorn SOP I.16
- Feeding frequency during acclimation: Detailed in STL Runcorn SOP I.16, with feeding stopped when juvenile turbot were transferred to test tanks 24 hours before test start
- Health during acclimation: Juvenile turbot were received on September 26, 2007 free of malformations and in good heath (Batch No. SM07002)

QUARANTINE
- Not applicable (Not wild caught)

FEEDING DURING TEST
- No

OTHER
- The test organism is the species recommended by the Paris Commission for studies on the effects of test chemicals on marine vertebrates

Test type:

semi-static

Water media type:

saltwater

Remarks:

Artificial seawater (Prodac Ocean Fish Marine Salt)

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

Observations were made at 0, 24, 48, 72 and 96 hours to determine toxicity after exposure to the test substance

Test temperature:

12.1 +/- 0.7 °C

pH:

7.92 +/- 0.2 in the control and 7.96 +/- 0.1 in treated media

Dissolved oxygen:

100.0 %

Salinity:

31.6 +/- 0.4 % in the control and 31.9 +/- 0.3 % in treated media

Nominal and measured concentrations:

Nominal concentrations of 0 (control) and 7.51 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: Not specified
- Material, size, headspace, fill volume: 10 litre volume of treated or control media
- Type of flow-through: Semi-static
- Renewal rate of test solution: Replacement of test and control media after 48 hours
- No. of organisms per vessel: 7 juvenile turbot
- No. of vessels per concentration: 1
- No. of vessels per control: 1
- No. of vessels per vehicle control: N/A

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Artificial seawater (Prodac Ocean Fish Marine Salt)
- Alkalinity: 7.72 - 8.21 pH
- Culture medium different from test medium: Animals were maintained as detailed in STL Runcorn SOP I.16 and held in artificial seawater (STL Runcorn SOP III.12). The test tanks contained artificial seawater of Prodac Ocean Fish Marine Salt
- Intervals of water quality measurement: Dissolved oxygen, pH, temperature, and salinity were recorded at 24 hour intervals

EFFECT PARAMETERS MEASURED: Mortality, i.e. cessation of movement and failure to respond to tactile stimulation. Observations were made at 0, 24, 48, 72 and 96 hours

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: A test was performed using the EC50 observed during the toxicity testing on algae to determine if fish is more sensitive to the test substance than Skeletonema costatum.
- Test concentrations: 0 (control) and 7.51 mg/L of p-toluene sulphonamido caproic acid morpholine salt

Reference substance (positive control):

not specified

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 7.51 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0% mortality

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 7.51 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0% mortality

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 7.51 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0% mortality

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 7.51 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 0% mortality

Sublethal observations / clinical signs:

After 96 hours exposure to p-toluene sulphonamido caproic acid morpholine salt, LC50 and percentage mortality in juvenile turbot were determined to be >7.51 mg/L and 0% respectively.

Mortality data for juvenile turbot exposed to control and test media containing p-toluene sulphonamido caproic acid morpholine salt:

Test Material Concentration (mg/L)		Exposure Period (hours)					Mortality at Test End (%)
		0	24	48	72	96
0	Alive	7	7	7	7	7	-
	Dead	0	0	0	0	0	0
7.51	Alive	7	7	7	7	7	-
	Dead	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

Following a short-term (96 hour) exposure of juvenile turbot to p-toluene sulphonamido caproic acid morpholine salt in artificial seawater, the LC50 and percentage mortality of the substance was reported as >7.51 mg/L and 0 %, respectively.

Executive summary:

The acute toxicity of p-toluene sulphonamido caproic acid morpholine salt to the fish Scophthalmus maximus (juvenile turbot) was determined in accordance to the OECD Guideline 203 (Fish, Acute Toxicty Test), with no reported deviation. The test substance was dispersed within artificial seawater media at a nominal concentration of 0 (control) and 7.51 mg/L and monitored under semi-static conditions for 96 hours. After 24, 48, 72, and 96 hours of exposure to p-toluene sulphonamido caproic acid morpholine salt, the percentage mortality in the treated vessel was found to be 0 % and an LC50 of >7.51 mg/L was subsequently reported. Similarly, all juvenile turbot in the control vessel remained mobile and responded to tactile stimulation by the test end.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 7 July 2008 to 11 July 2008

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:

other: OSPAR / PARCOM Protocols on Methods for the Testing of Chemicals Used in the Offshore Industry 2006

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A nominal 1,000 mg/L solution was prepared in dilulation water, shaken vigorously and allowed to stand for one hour.
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): 0 and 15 mg/L (limit test)
- Evidence of undissolved material: A clear colourless solution was obtained.

Test organisms (species):

Cyprinodon variegatus

Details on test organisms:

TEST ORGANISM
- Common name: Sheepshead Minnow
- Source: Opus Plus Ltd, Flotta, Stromness
- Length at study initiation: 17.7 mm (mean)
- Weight at study initiation: 0.07 g (mean)

ACCLIMATION
- Acclimation period: 5 days
- Acclimation conditions: same as test
- Type and amount of food during acclimation: Animals were fed daily using Tetramin Marine Flake Food at a rate of approximately 2% body weight per day until 24 hours before test initiation.
- Health during acclimation: No mortality observed

Test type:

semi-static

Water media type:

saltwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

21.5 to 22.5°C

pH:

7.9 - 8.1

Dissolved oxygen:

95 to 100% ASV

Salinity:

34%

Nominal and measured concentrations:

0 and 15 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: 12 L volume aquaria
- Type: open
- Renewal rate of test solution: Test solutions replaced after 48 hours.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): One replicate
- No. of vessels per control (replicates): One replicate

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

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Test concentrations: 0 and 15 mg/L
- Results used to determine the conditions for the definitive study: Results from previous short-term toxicity testing on algae and aquatic invertebrates

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: None observed
- Observations on body length and weight: No abnormalities reported
- Mortality of control: None observed
- Other adverse effects control: None reported
- Abnormal responses: None reported

Validity criteria fulfilled:

yes

Conclusions:

Following a short-term (96 hour) exposure of Sheepshead Minnow to p-toluene sulphonamidocaproic acid morpholine salt in artificial seawater, the LC50 and percentage mortality of the substance was reported as > 15 mg/L and 0 %, respectively.

Executive summary:

The acute toxicity of p-toluene sulphonamidocaproic acid morpholine salt to the fish Cyprinodon variegatus (Sheepshead Minnow) was determined in accordance to the OECD Guideline 203 (Fish, Acute Toxicty Test), with no reported deviation. The test substance was dispersed within artificial seawater media at a nominal concentration of 0 (control) and 15 mg/L and monitored under semi-static conditions for 96 hours. After 24, 48, 72, and 96 hours of exposure to p-toluene sulphonamidocaproic acid morpholine salt, the percentage mortality in the treated vessel was found to be 0% and an LC50 > 15 mg/L was subsequently reported. No abnormal behaviour was reported.

Description of key information

The acute toxicity of p-toluene sulphonamido caproic acid morpholine salt to marine water fish was determined in two studies conducted in accordance with the OECD Guideline 203 (Fish, Acute Toxicty Test), with no reported deviation.

Both studies were limit tests using the lowest EC₅₀/LC₅₀ determined in the short-term toxicity test to algae and aquatic invertebrates. The highest concentration tested was 7.51 mg/L during the first study and 15 mg/L during the second study.

No animal died or displayed abnormal behaviour as a result to the exposure to the test substance at both 7.51 mg/L and 15 mg/L.

Based on these results it is concluded that the LC₅₀ for the registered substance should be reported as > 15 mg/L.

Key value for chemical safety assessment

Marine water fish

Marine water fish

Effect concentration:

15 mg/L