4-morpholinopropanesulphonic acid

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8 October - 24 November 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

: The test temperature in the tests laid above the limit of 22 °C (maximum 23,8 °C). As the results of the study showed no inconsistencies, this deviation was stated as uncritical.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

yes

Remarks:

: The test temperature in the tests laid above the limit of 22 °C (maximum 23,8 °C). As the results of the study showed no inconsistencies, this deviation was stated as uncritical.

GLP compliance:

yes (incl. QA statement)

Remarks:

(Landesamt für Umweltschutz und Gewerbeaufsicht , Rheinallee 97-101 , D-55118 Mainz , Germany)

Test material

Test material information

Constituent 1

Reference substance name:

4-morpholinopropanesulphonic acid

EC Number:

214-478-5

EC Name:

4-morpholinopropanesulphonic acid

Cas Number:

1132-61-2

Molecular formula:

C7H15NO4S

IUPAC Name:

3-morpholin-4-ylpropane-1-sulfonic acid

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report) : MOPS HIGH PURITY
- Composition of test material : 3-(N-Morpholino)-propane sulfonic acid
- Substance type : pure substance
- Physical state : white powder
- Analytical purity : 100,3%
- Purity test date : not stated
- Lot/batch No. : 1190567/006
- Expiration date of the lot/batch : 31 August 2004
- Stability under test conditions : At the end of the test, the concentration of the test item in the treatment was determined as 105 % of the measured concentration at the beginning of the test. Therefore the test item can be stated as stable under test conditions.
- Storage condition of test material : The test item was stored in a closed polyethylene vessel at room temperature and kept away from ignition sources.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The real concentration of the test item in the treatment was determined before and after the test.

Test solutions

Vehicle:

no

Details on test solutions:

Pre-Test
Preparation of the test solutions :
A stock solution containing 1000 mg/L in dilution water was prepared . The stock solution then was used to prepare four concentrations in a geometric series , spaced by the factor 10.

Main Study :
Preparation of the test solutions
The result of the pre-test showed that the EC50 must lie above 1000 mg/L . Therefore the main study was performed as a limit test. A solution containing 1000 mg/L was prepared.

Reference Test :
Preparation of the test solutions
A stock solution containing 250 mg K2Cr2O7 /L in deionized water was diluted with dilution water to give a series of seven concentrations between 0,625 and 3,0 mg/L.

Dilution Water
Drinking water with an enrichment of certain minerals was used in the test.
Amount added:
CaCl2*2H2O : 179,0 mg/L
MgSO4*7H2O : 69,5 mg/L
NaCl : 16,1 mg/L
KCl : 0,65 mg/L
After preparation, the dilution water was aerated, and the pH was adjusted to 7,8 ± 0,2.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name : Water flea
- Source : Origin : Umweltbundesamt Berlin
- Age at study initiation (mean and range, SD): between 0 and 24 h
- Method of breeding: maintained in glass beakers with a nominal volume 2 L containing M4-Medium (see "Any other information on materials and methods incl. tables") at temperature 20±2°C; 16 hours light, 8 hours dark , using neon tubes ; Food : cold water fish food Tetra Aminin®, finely ground, suspended in M4-Medium and boiled for three seconds, green algae (Desmodesmus subspicatus) ; changing of medium : twice a week
- Feeding during test : none

Selection of Daphnia
24 hours before the start of the test, the adult animals were separated from the youngs . 23 hours later, the adults were caught with the help of a glass tube, and the newborn daphnia , aged between 0 and 23 hours, were sieved from the medium and immediately placed into a 250 ml-beaker containing dilution water. After a settling-in period of 30 minutes, animals which showed no apparent damage were used for the test.
There was no difference in the selection of animals in pre-test and main study.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

no data

Test temperature:

20 ± 2°C (deviation within the test ± 1°C)

pH:

Pre-Test : 6,6 - 8,1
Main Study : 6,8 - 6,9
Reference Test : 7,3 - 7,8

Dissolved oxygen:

Pre-Test : 8,0 - 8,3 mg/L
Main Study : 7,5 - 8,5 mg/L
Reference Test : 7,5 - 8,7 mg/L

Nominal and measured concentrations:

The analytical determination of the test item in the test solution showed a very good recovery after 48 hours.
The recovery of more than 100 % was probably caused by evaporation effects. The test item can be considered as stable under the test conditions.
(For Details see "Any other information on materials and methods incl. tables")

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type : open
- Material, size, headspace, fill volume : glass beakers , nominal volume 50 ml containing 20 ml test solution , tall shape
- Aeration : none
- Renewal rate of test solution (frequency/flow rate) : none
- No. of organisms per vessel : 5
- No. of vessels per concentration (replicates) : 2 in the pre-test ; 4 in the main study
- No. of vessels per control (replicates) : 4
- No. of vessels per blank (replicates) : 2 in the pre-test ; 4 in the main study

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water : Drinking water with an enrichment of certain minerals was used in the test.
Amount added:
CaCl2*2H2O : 179,0 mg/L
MgSO4*7H2O : 69,5 mg/L
NaCl : 16,1 mg/L
KCl : 0,65 mg/L
After preparation, the dilution water was aerated, and the pH was adjusted to 7,8 ± 0,2.


OTHER TEST CONDITIONS
- Lighting : none
- Feeding : none

TEST CONCENTRATIONS
- Pre-Test : 1000/100/10/1 mg/L
- Main study : 1000 mg/L
- Results used to determine the conditions for the definitive study : The result of the pre-test showed that the EC50 must lie above 1000 mg/L. Therefore the main study was performed as a limit test. A solution containing 1000 mg/L was prepared.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate K2Cr2O7 (CAS No. 7778-50-9)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

other: NOEL

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilization

Remarks on result:

other: not determined

Effect concentrations 2

Duration:

24 h

Dose descriptor:

other: EC50i

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilization

Remarks on result:

other: not determined

Effect concentrations 3

Duration:

24 h

Dose descriptor:

other: EC100i

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilization

Remarks on result:

other: not determined

Effect concentrations 4

Duration:

48 h

Dose descriptor:

other: NOEL

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilization

Remarks on result:

other: not determined

Effect concentrations 5

Duration:

48 h

Dose descriptor:

other: EC50i

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilization

Remarks on result:

other: not determined

Effect concentrations 6

Duration:

48 h

Dose descriptor:

other: EC100i

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: immobilization

Remarks on result:

other: not determined

Details on results:

In the initial experiment, four concentrations of the test item in a geometric series (spaced by factor 10, the highest being 1000 mg/L) were used. For each concentration, ten daphnia were exposed to the test item for 48 hours.
No concentration caused immobilization towards the daphnia.
None of the animals showed signs of morbidity.

The main study was performed as a limit test using the nominal concentration 1000 mg/L, which showed no effect on the daphnia. The real concentration of the test item in the treatment was determined before and after the test. The test item can be stated as stable under the test conditions. Therefore the nominal concentration was used to estimate the results.

Results with reference substance (positive control):

For the estimation of the EC50 of the reference substance, the fit showed sufficient statistical correspondance of the data with the dose-response-equation.

Any other information on results incl. tables

Findings

Pre-Test

Conc. in mg/L	Immobility abs. t=24 +48h		Immobility in % t=24 +48h	pH (t=0h)	pH (t=48h)	O2 -conc. mg/L (t=0h)	O2 -conc. mg/L (t=48h)
0	0	0	0	7,7	8,1	8,3	8,1
1000	0	0	0	6,6	7,0	8,3	8,1
100	0	0	0	7,4	8,1	8,3	8,1
10	0	0	0	7,8	8,1	8,3	8,0
1	0	0	0	7,8	8,0	8,3	8,0

Main Study

Conc. in mg/L	Immobility abs. t=24 +48h				Immobility in % t=24 +48h	pH (t=0h)	pH (t=48h)	O2-conc. mg/L (t=0h)	O2-conc. mg/L (t=48h)
0	0	0	0	0	0	8,1	7,9	8,5	7,8
1000	0	0	0	0	0	6,9	6,8	8,5	7,5

Reference Test

Conc. in mg/L	Immobility abs.				Immobility in %	pH (t=0h)	pH (t=48h)	O2-conc. mg/L (t=0h)	O2-conc. mg/L (t=48h)
0	0	0	0	0	0	7,7	7,7	8,6	7,8
3,0	5	5	5	5	100	7,8	7,7	8,6	7,8
2,5	5	5	5	5	100	7,8	7,6	8,7	7,8
2,0	5	4	5	3	85	7,8	7,7	8,6	7,8
1,5	3	2	4	1	50	7,8	7,6	8,6	7,7
1,25	1	0	0	0	5	7,8	7,8	8,6	7,7
1,0	0	0	0	0	0	7,7	7,7	8,6	7,6
0,625	0	0	0	0	0	7,7	7,3	8,5	7,5

Validity criteria

- The 24h EC50i of K₂Cr₂O₇ should lie between 0,6 and 2,4 mg/L. The 24h EC50i of K₂Cr₂O7 was determined as 1,5 mg/L.

- Immobilization in the blanks may not exceed 10 %. Immobilization in the blanks was 0 %.

- The concentration of dissolved oxygen at the end of the test must be at least 60% of the saturation value . The lowest concentration of dissolved oxygen at the end of the test was 7,5 mg/L. 100% saturation equals 9,05 mg/L at 20 °C, giving 83 % oxygen saturation for the value measured.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

all validity criteria fulfilled

Conclusions:

The study is regarded as a valid guideline study with certificated GLP compliance. According to the reported EC50 value (> 1000 mg/L) the test substance is not classified as toxic to aquatic invertebrates.

Executive summary:

The acute toxicity to aquatic invertebrates of the test substance was tested according to OECD Guideline 202 (EU Method C.2) in a static freshwater test with Daphnia magna as test organisms. The study was conducted as a limit test under certificated GLP compliance. The test duration was 48 hours with a substance concentration of 1000 mg/L, which was selected based on the preliminary range-finding study. Potassium dichromate was used as reference substance. The validity criteria were met. At the end of the test, the concentration of the test item in the treatment was determined as 105 % of the measured concentration at the beginning of the test. Therefore the test item can be stated as stable under test conditions. For this reason, the results are given in nominal concentrations. The following deviation from the study plan was documented and assessed : The test temperature in the tests laid above the limit of 22°C (maximum 23,8°C). As the results of the study showed no inconsistencies, this deviation was stated as uncritical. The following results were determined for the test item MOPS HIGH PURITY in the acute toxicity test against Daphnia magna :

- 24h EC50i : > 1000 mg/L

- 24h EC100i : > 1000 mg/L

- 24h NOEL : 1000 mg/L

- 48h EC50i : > 1000 mg/L

- 48h EC100i : > 1000 mg/L

- 48h NOEL : 1000 mg/L