3,3'-methylenebis[5-methyloxazolidine]

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:

2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Study according to guideline (OECD 202, Part I) in compliance with GLP. The reported pH was above 9 (pH 9.1, 9.3 and 9.4 at 10, 80 and 100 mg/L, respectively) after application. Only for these three concentration levels immobilisation was reported. After 48 hours reported pH values were all below 9, however mortality reached already 100% for 80 and 100 mg/L and 75% for 40 mg/L after 24 hours.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Report date:

2000

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Institut Fresenius

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: Lot No. 645109701
- Specification: 3,3’- Methylene-bis [5-methyl-oxazolidine], 50-100%
- Colourless liquid with a typical odour
- Purity: > 95%

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Nominal concentrations 20, 40 and 80 mg/l were measured as fresh solution and after 72 h.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

- Strain: Daphnia magna STRAUS
- Source: Laboratory bred, originally derived from Dr. Knie, LWA Düsseldorf (Germany)
- Age: ≤ 24 h
- Feeding of animals during test: No

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

20.5 – 22.0 °C

pH:

7.31 – 9.38

Dissolved oxygen:

7.4 – 8.0 mg/L

Nominal and measured concentrations:

- Nominal concentrations: 100, 80, 40, 20, and 10 mg/L + control
- Measured concentrations: Nominal concentrations 20, 40, and 80 mg/l analyzed as fresh solution and after 72 h (For values see table below)

Details on test conditions:

TEST MEDIUM / WATER PARAMETER
- dilution water was prepared according to DIN 38412, part 11

TEST SYSTEM
- Static system / No renewal of test solution
- Volume of test vessels: 290 mL
- Volume/animal: 58 mL
- Number of animals/vessel: 5
- Number of vessels/ concentration: 4
- Test performed in closed vessels due to significant volatility of TS: Test solutions filled into the carboys avoiding any space of air, stoppered with glass stoppers

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Aeration of dilution water: No
- Quality/Intensity of irradiation: No data
- Photoperiod: Light/dark cycle 16 h / 8 h

TEST PARAMETER
- Immobility (no swimming within 15 sec.; moving of antenna was not considered as swimming).

SAMPLING
- Immobilisation recorded after 48 h
- pH, O2-content and temperature measured at t(0) and after 48 h

STATISTICS
- EC50 determined as geometric mean between NOEC and EC100

Reference substance (positive control):

no

Remarks:

No reference substance applied during this test. However, sensitivity of test system towards K-dichromate was tested once a month.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

40 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

57 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

80 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

11 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

29 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 6

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

75 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- The EC0 and EC100 were directly taken from the concentrations being tested. EC50 is the geometric mean between EC0 and EC100.
- No other effects were observed.

Results with reference substance (positive control):

EC50 = 1.2 mg/L during the test period.

Any other information on results incl. tables

Immobility data of exposed daphnids:

 

Test substance Concentration (nominal/) [mg/l]
	ImmobileDaphnia
	Number		Percentage		O2 [mg/l] 48 h	pH   48 h	Temp. [°C] 48 h
	24 h	48 h	24 h	48 h
Control	No data		0	0	7.5	7.40	20.5
100			100	100	7.7	9.02	20.5
80			100	100	7.5	8.92	20.5
40			75	100	7.5	8.79	20.5
20			0	0	7.4	8.57	20.5
10			0	0	7.4	8.38	20.5

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test results indicate that the substance is acutely harmful towards invertebrates.

Executive summary:

The substance was found to be acutely toxic towards daphnids. All validity criteria according the test Guideline are met within the study.

A steep concentration-response relationship can be established from the study results. Exposure at 40 mg/L resulted in 100% mortality within 48 hours. At 20 mg/L no mortality was observed until test termination. The EC50 was calculated as the geometric mean between EC0 and EC100.

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 thethe low adsorptivity of the test substance  influenced test substance concentrations. Possible substance losses due to biodegradation are unlikely during the short test period of 48 h.

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 free formaldehyde content revealed that the deviation from nominal values were <20%. Therefore, the nominal concentrations can be considered as effective concentrations.