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

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key 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:

other: ISO 14669:1999 (E) Water Quality - Determination of Acute Lethal Toxicity to Marine Copepods (Copepoda, Crustacea)

Version / remarks:

Draft method

Deviations:

no

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 was initially prepared at a 2.0 g/L stock solution in 25 μm filtered seawater, as the test material was described as being soluble in water. Aliquots of the stock solution were further diluted to achieve the appropriate range of test concentrations
- Control: The control, 25 μm filtered seawater, was prepared as a bulk volume of 50 mL from which 5 mL volumes were removed and added to appropriately labelled vessels. A reference material, 3,5-dichlotophenol at a concentration of 1.0 mg/L, was also prepared in filtered seawater and added to labelled vessels
- Evidence of undissolved material: No

Test organisms (species):

other aquatic crustacea: Acartia tonsa

Details on test organisms:

TEST ORGANISM
- Common name: Acartia tonsa
- Justification for species other than prescribed by test guideline: Acartia tonsa was selected as a suitable representative of marine herbivores, which is accepted under the UK Harmonised Offshore Chemical Notification Scheme (HOCNS) and by the Paris Commission (PARCOM) signatory nations
- Source: Guernsey Sea Farms, Guernsey
- Feeding: Not specified, all animals were maintained as detailed in STL Runcorn SOP I.7 (Batch No. AT07024)

QUARANTINE
- Not applicable (not wild caught)

OTHER
- At the start of the test the Acartia tonsa nauplii were 13-15 days old

Test type:

static

Water media type:

saltwater

Remarks:

25 μm filtered seawater

Limit test:

no

Total exposure duration:

48 h

Remarks on exposure duration:

Observations were noted at 24 and 48 hours of exposure to the test material

Test temperature:

Temperature was recorded at the beginning of the study and study-end. Seawater temperature on arrival was reported at 17 °C and prior to use the test media was stored at 20 ± 2 °C

pH:

pH was recorded at the beginning of the study and study-end. The pH of the seawater on arrival was 8.08

Dissolved oxygen:

Dissolved oxygen was recorded at the beginning and end of the study. The dissolved oxygen level of the seawater was 100% on arrival

Salinity:

An arrival seawater salinity was reported to be 33 g/L

Nominal and measured concentrations:

From the 2.0 g/L stock solution, nominal concentrations of p-toluene sulphonamido caproic acid morpholine salt of 112.5, 200.0, 355.6, 632.5, 1124.7, and 2000.0 mg/L were prepared. Vessels of 3,5-dichlorophenol at a concentration of 1.0 mg/L were formulated in addition

Details on test conditions:

TEST SYSTEM
- Information not specified. The study was undertaken according to the draft method of ISO 14669:1999 (E) and as detailed in STL Runcorn SOP III.14

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Seawater was filtered to 25 μm
- Water parameters: Alkalinity (pH), salinity, and temperature was 8.08, 33.6 g/L, and 20.0 °C on arrival, respectively, with seawater stored at 20 ± 2 °C prior to use. Additional information was not specified
- Intervals of water quality measurement: Temperature, pH, salinty, and the dissolved oxygen level were measured at the beginning at end of the study, however, this data was not published in the report

EFFECT PARAMETERS MEASURED: Mortality of Acartia tonsa was noted at 24 and 48 hours in order to determine the median effect concentration that kills 50% of animals exposed (LC50), 95% fiducial limits, No Observed Effect Concentration (NOEC), and Lowest Observed Effect Concentration (LOEC) using the statistical computer programme Toxcalc (Tidepool Scientific, STL Runcorn SOP V.2.)


RANGE-FINDING STUDY
- Test concentrations: On receipt of p-toluene sulphonamido caproic acid morpholine salt, a range-finder study was undertaken using 0, 2.0, 20.0, and 200.0 mg/L
- Results used to determine the conditions for the definitive study: A definitive test range of 112.5 - 2000.0 mg/L was defined (see results table)

Reference substance (positive control):

yes

Remarks:

3,5-Dichlorophenol (1.0 mg/L)

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

572.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

322 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

632.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

1 124.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

- Mortality of control: Immobilisation reported as 7 and 13 % at 24 and 48 hours in the seawater control (0 mg/L), respectively, and 20% at 24 and 48 hours in the 3,5-dichlorophenol (1.0 mg/L) positive control

Results with reference substance (positive control):

- Results with reference substance valid: Yes
- Relevant effect levels: Yes, immobilisation of 20% of Acartia tonsa had occurred at 24 and 48 hours in the test media containing 3,5-dichlorophenol

Reported statistics and error estimates:

Observation data regarding mortality was processed for each time point with the statistical computer programme Toxcalc, supplied by Tidepool Scientific (STL Runcorn SOP V.2.). Toxcalc was used to determine the substance LC50, 95% fiducial limits, NOEC, and LOEC

Percentage immobilisation data (%) for the copepod Acartia tonsa exposed to p-toluene sulphonamido caproic acid morpholine salt in a range-finder study:

Test material concentration (mg/L)	Immobilisation after 48 hours (%)
0	7
2.0	20
20.0	10
200.0	40

From the observed results, a definitive range of 112.5 - 2000.0 mg/L was defined

Percentage immobilisation data (%) for the copepod Acartia tonsa exposed to p-toluene sulphonamido caproic acid morpholine salt:

Test material concentration (mg l-1)	Immobilisation (%)
	24 hours	48 hours
0.0	7	13
112.5	0	0
200.0	35	50
355.6	35	55
632.5	25	80
1124.7	80	100
2000.0	100	100
3,5-dichlorophenol	20	20

Validity criteria fulfilled:

yes

Remarks:

All parameters measured in this study lay within acceptable limits of tolerance

Conclusions:

Acartia tonsa was exposed to p-toluene sulphonamido caproic acid morpholine salt at a definitive range of 112.5 - 2000.0 mg/L. After 24 and 48 hours, the test substance was determined to have an EC50 of 572.7 and 322.0 mg/L, respectively. In addition, the highest concentration of test material producing No Observable Effect (NOEC) was 632.5 mg/L, while the Lowest Observed Effect Concentration (LOEC) in the study was reported to be 1124.7 mg/L.

Executive summary:

A test carried out in accordance with draft method ISO 14669:1999 (E) Water Quality - Determination of Acute Lethal Toxicity to Marine Copepods (Copepoda, Crustacea) was used to determine the acute toxicity potential of p-toluene sulphonamido caproic acid morpholine salt, without no recorded deviations from the test method. To determine the appropriate dose level of the test substance a range-finding test at 0 - 200.0 mg/L was performed, with the following nominal concentrations having been selected for exposure to Acartia tonsa: 0 (control seawater), 112.5, 200.0, 355.6, 632.5, 1124.7, and 2000.0 mg/L. 3,5 -dichlorophenol at a concentration of 1.0 mg/L was used as a positive control reference substance within filtered seawater. There was clear evidence of dose response in this study. After 24 hours of exposure to p-toluene sulphonamido caproic acid morpholine salt, the concentration at which 50% of Acartia tonsa were killed was calculated to be 572.7 mg/L, and at 48 hours was 322.0 mg/L. The No Observed Effect Concentration (NOEC) and lowest concentration producing 100% mortality (LOEC) were determined to be 632.5 mg/L and 1124.7 mg/L, respectively. The only treatment in the study, including the control, that did not elicit mortality was 112.5 mg/L.

Description of key information

A test carried out in accordance with draft method ISO 14669:1999 (E) Water Quality - Determination of Acute Lethal Toxicity to Marine Copepods (Copepoda, Crustacea) was used to determine the acute toxicity potential of p-toluene sulphonamido caproic acid morpholine salt, without no recorded deviations from the test method. To determine the appropriate dose level of the test substance a range-finding test at 0 - 200.0 mg/L was performed, with the following nominal concentrations having been selected for exposure to Acartia tonsa: 0 (control seawater), 112.5, 200.0, 355.6, 632.5, 1124.7, and 2000.0 mg/L. 3,5 -dichlorophenol at a concentration of 1.0 mg/L was used as a positive control reference substance within filtered seawater. There was clear evidence of dose response in this study. After 24 hours of exposure to p-toluene sulphonamido caproic acid morpholine salt, the concentration at which 50% of Acartia tonsa were killed was calculated to be 572.7 mg/L, and at 48 hours was 322.0 mg/L. The No Observed Effect Concentration (NOEC) and lowest concentration producing 100% mortality (LOEC) were determined to be 632.5 mg/L and 1124.7 mg/L, respectively. The only treatment in the study, including the control, that did not elicit mortality was 112.5 mg/L.

Key value for chemical safety assessment

Marine water invertebrates

Marine water invertebrates

Effect concentration:

322 mg/L