3,3'-methylenebis[5-methyloxazolidine]

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Study according to guideline (OECD 201, EG C.3) in compliance with GLP. In the vessels with the highest applied test substance concentration (23.43 mg/L) the pH value deviated by more than 1 unit during the test. This deviation is considered to have no influence on the integrity of the results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Remarks:

In the vessels with the highest applied test substance concentration (23.43 mg/L) the pH value deviated by more than 1 unit during the test.

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

GLP compliance:

yes (incl. QA statement)

Remarks:

Arbeitsgemeinschaft GAB/IFU GmbH

Test material

Test material information

Constituent 1

Reference substance name:

3,3'-methylenebis[5-methyloxazolidine]

EC Number:

266-235-8

EC Name:

3,3'-methylenebis[5-methyloxazolidine]

Cas Number:

66204-44-2

Molecular formula:

C9H18N2O2

IUPAC Name:

3,3'-methylenebis[5-methyloxazolidine]

Details on test material:

- Test material: Reaction products of paraformaldehyde and 2-hydroxypropylamine (ratio of 3:2)
- Lot/Batch number: Batch number: NA 2571
- Specification: MAR 71
- Clear liquid; infinitely soluble in water; density ca. 1.06 kg/L
- Purity: 95% (w/w)
- Further relevant properties: vapour pressure < 1 hPa

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Nominal concentrations 4.8 and 23.4 mg/l were measured as fresh solution and after 24, 48, and 72 h.

Test organisms

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

- Species: Scenedesmus subspicatus CHODAT (current name Desmodesmus subspicatus)
- Strain: No data
- Source: Origin: Pflanzenphysiologisches Institut der Universität Göttingen (Germany)
- Laboratory culture: Yes
- Method of cultivation: Stock cultures were continuously grown in aerated semi-static liquid cultures. Stock cultures also kept on slant agar tubes and freshly passaged about once a month.
- Pretreatment: No data
- Initial cell concentration: 104 cells/mL

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Test temperature:

21.0 – 25.0 °C

pH:

7.96 – 9.22

Nominal and measured concentrations:

Nominal concentrations:
- Range-finding test: 0.01, 0.1, 1, 10, 100 mg/L + control
- Main test: 1.0, 2.2, 4.84, 10.65, 23.43 mg/L + control (nominal concentrations)

Measured concentrations:
- Test substance measured in assays with 0, 4.84 and 23.43 mg/L at different timepoints without inoculation (For values see table below)

Details on test conditions:

GROWTH MEDIUM
- Standard medium used: yes (OECD TG 201 medium)

TEST SYSTEM
- Volume of culture flasks: 1000 mL Duran bottles, 500 mL test volume
- Light quality: 2 light tubes (OSRAM L 36W/25, DIN IEC 81, light colour 25, universal white)
- Number of vessels/ concentration: 3 (test substance); 6 (control)

OTHER TEST CONDITIONS
- Aeration of dilution water: The algae were supplied with CO2 by aeration. Air was filtered through activated carbon and sterile membrane filters.

SAMPLING
- Cells counted after 24, 48 and 72 h
- pH measured at beginning and end of test. Temperature measured after 0, 24, 48 and 72 h.

TEST PARAMETER
- Growth rate, biomass

STATISTICS
- Mean value for each MAR 71 concentration and control plotted versus time. EbC, ErC and NOEC values calculated by log-linear regression analysis.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

2.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

2.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

5.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 4

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 5

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

1.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 6

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

2.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

- No other effects obseved

Any other information on results incl. tables

Results of main test:

 

Test substance Concentration (nominal) [mg/l]	Cell concentrations (mean values) [10^4cells/ml]
	measured				Inhibition Percent
	0 h	24 h	48 h	72 h	biomass	growth
control	1.0	5.73	22.92	121.6
1.0	1.0	3.13	25.52	144.3	-13.0	-3.8
2.2	1.0	2.61	15.10	81.25	35.8	9.0
4.84	1.0	2.60	2.61	12.50	89.7	48.3
10.65	1.0	0.00	1.56	3.65	99.0	73.3
23.43	1.0	0.00	1.56	0.52	100.8	96.9
Temp.[°C]	21.0-25.0	24.0-25.0	23.0-25.0	23.0-25.0
pH	7.96-9.22			7.96-9.15

 

Cell concentrations are mean of 3 parallels.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The substance was found to be harmful towards algae at relatively low concentrations.

Executive summary:

The substancewas found to be acutely toxic towards algae at relatively low concentrations.The numerical results are given below; all results refer to the nominal concentrations.All validity criteria according the test Guideline are met within the study.

A clear concentration-response relationship can be established from the study results.

The constituents of the reaction product (substance) are not volatile from aqueous solutions, therefore this removal mechanism could not influence the test results. It is not expected that the the low adsorptivity of the test substance influenced test substance concentrations.

The constituents of the reaction product (substance) hydrolyse completely in concentrations which are expected to occur in waste waters and surface waters. Also in the media of toxicity tests, complete hydrolysis is expected, therefore the observed effects are assumed to be caused by a mixture of hydrolysis products.

Analytical measurements of the formaldehyde content in the test substance was not successful. During the test period an increase of aldehyde groups was observed which was probably caused by injection of aldehydes via the aeration system. Measurements in test solutions of the tests to daphnia and fish revealed that the test substance was stable during the test period. Therefore, in the present test the effective concentrations can be considered to be equal to the nominal concentrations.

In the vessels with the highest applied test substance concentration (23.43 mg/L) the pH value deviated by more than 1 unit during the test, but not in the vessels with lower concentrations. Therefore, this deviation should not influence the calculated effect values.