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Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
13 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 guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
yes
Remarks:
:For each study a stock solution of an appr. conconcentration, and not 1 g/L as stated in the study plan, was prepared.Test temperature after 24h of the pre-test laid above the limit of 24°C(max. 24,2°C).As all these deviations were stated as uncritical .
Qualifier:
according to guideline
Guideline:
EU Method C.1 (Acute Toxicity for Fish)
Deviations:
yes
Remarks:
: For each study a stock solution of an appr. conconcentration, and not 1 g/L as stated in the study plan, was prepared.Test temperature after 24h of the pre-test laid above the limit of 24°C(max. 24,2°C).As all these deviations were stated as uncritical
GLP compliance:
yes (incl. QA statement)
Remarks:
(Landesamt für Umweltschutz und Gewerbeaufsicht , Rheinallee 97-101 , D-55118 Mainz , Germany)

Test material

Constituent 1
Chemical structure
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
- Physical state : white powder
- Analytical purity : 100,3%
- Composition of test material : 3-(N-Morpholino)-propane sulfonic acid
- Lot/batch No. : 1190567/006
- Expiration date of the lot/batch : 2004-08-31
- Stability under test conditions : The analytical determinations of the test item in the test solution showed good recovery after 96 hours. Therefore the test item can be considered as stable under the test conditions.
- Storage condition of test material : The test item was stored in a closed polyethylene vessel at room temperature

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
- Sampling method : Samples were taken at start and at the end of the test

Test solutions

Vehicle:
no
Details on test solutions:
For each study, a stock solution of an appropriate concentration in drinking water was prepared. The stock solution then was used to prepare solutions of the concentrations to be tested.
Pre-Test
Date : Oct. 13th – Oct 17th 2003
Stock solution : 6600 mg/L
Treatments : 1000 / 100 mg/L
Duration : 96 hours
Three fish were used for each concentration and for the blank.

Main Study
Date : Nov. 18th – Nov. 22nd 2003
Stock solution : 10000 mg/L
Treatment : 1000 mg/L
Duration : 96 hours
Seven fish were used for the test solution and for the blank.

Test organisms

Test organisms (species):
other: Danio rerio , Hamilton-Buchanan
Details on test organisms:
TEST ORGANISM
- Common name : zebrafish
- Source : Weiler's Pet's best, Turmstr. 6, D-67433 Neustadt, Germany
- Age at study initiation : sexually immature young fish
- Length at study initiation : 2 ± 1 cm
- Feeding during test : none

ACCLIMATION
- Acclimation period : 14 days
- Acclimation conditions : same as test
- Type and amount of food : type not stated , about 1-2 % of body weight per day
- Feeding frequency : three times a day
- Health during acclimation : During this period, mortality didn't surpass 5 %

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h

Test conditions

Hardness:
not stated
Test temperature:
23 ± 1 °C
pH:
6,7 - 7,9
Dissolved oxygen:
8,0 - 9,2 mgO2/L
Nominal and measured concentrations:
For the determination of the real concentrations, the area of the measured reference solution with 1 g test item/L was used (Area 2,231). The analytical determinations of the test item in the test solution showed good recovery after 96 hours. The recovery of more than 100 % was probably caused by evaporation effects. Therefore the test item can be considered as stable under the test conditions.
The measured concentrations of the test item are presented in table 1 at "any other information on results incl. tables".

Details on test conditions:
TEST SYSTEM
- Test vessel
- Material, size, headspace, fill volume : Glass aquaria , maximal volume 10 l
- Aeration : accomplished with glass tubes, frequency of bubbles 1/s
- No. of organisms per vessel : 7
- No. of vessels per concentration (replicates) : 1
- No. of vessels per control (replicates) : 1
Observations were made every 24 hours, measuring pH and 02-concentration of the test solution in each vessel and recording mortalities or abnormal behaviour.In the mainstudy, the real concentration of the test item in the solutions were determined at the start and at the end of the test.
A fish was considered dead, if no visible movement could be observed, and if touching of the caudal peduncle produced no reaction.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water : Good quality drinking water is used. The composition is analysed yearly by the supplier.
ANALYSIS OF DRINKING WATER
Date of analysis: March 2003 , Performed by: Technologiezentrum Wasser, Karlsruhe
pH : 8,37
pH following CaCO3 saturation : 8,12
conductivity : 251 μS/cm
aluminium : < 0,02 mg/L
ammonium : < 0,01 mg/L
bor :< 0,02 mg/L
calcium : 30,0 mg/L
chloride : 11,3 mg/L
iron : < 0,01 mg/L
potassium : 2,7 mg/L
nitrogen (Kjeldahl) : < 0,05 mg/L
magnesium : 5,0 mg/L
mangane : < 0,005 mg/L
sodium : 11,2 mg/L
phenols : < 0,01 μg/L
phosphor : < 0,01 mg/L
silver : < 0,0005 mg/L
sulphate : 17,4 mg/L
dissolved or emulgated hydrocarbons : < 0,01 mg/L
substances which are extractable with chloroform : < 0,01 mg/L
surfactants : < 0,05 mg/L
arsenic : < 0,001 mg/L
lead : < 0,001 mg/L
cadmium : < 0,0001 mg/L
chromium : < 0,001 mg/L
cyanide : < 0,01 mg/L
fluoride : < 0,05 mg/L
nickel : < 0,001 mg/L
nitrate : < 1,0 mg/L
nitrite : < 0,01 mg/L
mercury : < 0,05 μg/L
polycyclic aromatic hydrocarbons :m < 0,05 μg/L
chlorinated organic compounds : < 0,01 mg/L
pesticides : < 0,1 μg/L
antimony : < 0,001 mg/L
selenium : < 0,001 mg/L

OTHER TEST CONDITIONS
- Adjustment of pH : none
- Photoperiod : 12/12 hours using neon tubes
Reference substance (positive control):
no

Results and discussion

Effect concentrations
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality (fish)
Details on results:
- Behavioural abnormalities :
No abnormal behaviour of the surviving fish could be observed

- Other biological observations:
see Biological results at "any other information on results incl. tables".

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
The measured concentrations at the start of the test showed good correlation with the nominal concentrations (82 -104 %). At the end of the test, only 4-90 % of the nominal concentration were detected. The recovery rate after 96 hours was therefore only in a range of 4 and 86 %. As the test item was found as stable under the conditions of the test in a concentration of 100 mg/L and the absolute loss of the test item during the test was nearly the same in each test solution (ranging from 0.8 to 1.5 mg/L), it can be assumed that the reduction of the concentration was caused by an uptake through the fish. Therefore the geometric mean of the measured concentrations was used for the determination of the results.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1 Measured Concentrations :

Nom. Conc. in mg/L  Area resp. Real Conc. in mg/L           % recovery after 96 h 
  0 h     96 h      
  Area  Conc.  Area  Conc.   
-- 
1000  2,14  959,21  2,274  1019,27  106,26

Table 2 : Results

Parameter

VALUE

95%-confidence-interval

24 -96h LC0

1000 mg/L

n. a.

24 -96h LC50

> 1000 mg/L

n. a.

24 -96h LC10O

>> 1000 mg/L

n. a.

Findings Pre-Test

Mortalities

Conc. in mg/L

Total   of

fish used

dead   fish

t = 24 h

dead   fish

t= 48h

dead   fish

t = 72 h

 dead fish t = 96 h
blank 
1000 
100

No abnormal behaviour of the surviving fish could be observed .

pH and 02 concentrations

Conc.

pH

02 in mg/L

mg/L

0 h

24h

48h

72h

96h

0 h

24h

48h

72h

96h

blank

7,8

7,8

8,0

8,2

8,5

7,9

8,0

8,2

8,3

8,4

1000

7,9

7,6

7,5

7,3

7,2

8,0

8,0

8,3

8,4

8,4

100

7,8

7,9

8,0

8,3

8,4

7,9

8,0

8,1

8,5

8,3

Findings Main Study

Mortalities

Conc. in mg/L

Total   of

fish used

dead   fish

t = 24 h

dead   fish

t= 48h

dead   fish

t = 72 h

 dead fish t = 96 h
blank 
1000 

No abnormal behaviour of the surviving fish could be observed

pH and 02concentrations

Conc.

pH

02in mg/L

mg/L

0 h

24h

48h

72h

96h

0 h

24h

48h

72h

96h

blank

8,6

7,9

7,8

7,9

7,7

8,7

8,2

8,1

8,2

8,5

1000

6,8

7,8

7,9

6,7

6,8

9,2

7,9

8,0

8,2

8,5

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
The mortality in the control may not exceed 10% at the end of the test. No mortalities occurred in the control;The dissolved O2 concentration must be at least 60 % throughout the test.Concentration of dissolved oxygen stayed above 92% throughout the test
Conclusions:
The study is regarded as a valid guideline study with certificated GLP compliance. According to the reported LC50 value (> 1000 mg/L) the test substance is not classified as toxic for fish.
Executive summary:

The acute toxicity to fish of MOPS HIGH PURITY was tested according to OECD guideline 203 and EU Method C.1 in a static freshwater test with zebrafish (Danio rerio, Hamilton-Buchanan) as test organisms. The study was conducted under certificated GLP compliance. The validity criteria were met. In the initial experiment, two concentrations of the test item (1000 and 100 mg/L) were used. For each concentration, three fish were exposed to the test item for 96 hours. No concentration caused mortality towards the fish. None of the animals showed signs of morbidity at the end of the test. The pH and the oxygen value of the treatments were normal. The main study was performed as a limit test using the nominal concentration 1000 mg/L which showed no effect on the fish. 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 result.

The following result was determined for the test item MOPS HIGH PURITY acute toxicity test against Danio rerio, HAMILTON BUCHANAN :

96h-LC50 : > 1000 mg/L