3,3'-methylenebis[5-methyloxazolidine]

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Study according to guideline (OECD 203) in compliance with GLP.

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 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

The temperature range over the test period (range 20.0 – 22.9 °C) was slightly out of the requested (within 2°C).

GLP compliance:

yes (incl. QA statement)

Remarks:

Istitut 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 25, 50 and 100 mg/l were measured as fresh solution and after 24 h.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

- Source: Tagis-Aquarium, Dreieich (Germany), and Aquafarm Ryba, Zeven (Germany)
- Wild caught: No
- Age/size: Length ca. 3 cm
- Feeding of animals during test: No

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Test temperature:

20.0 – 22.9 °C

pH:

7.52 – 8.44

Dissolved oxygen:

7.2 – 8.1 mg/L (Aeration of dilution water during test)

Nominal and measured concentrations:

Nominal concentrations: 100, 50, 25, 12.5, and 6.25 mg/L + control
Measured concentrations: Nominal concentrations 25, 50, and 100 mg/l analyzed as fresh solution and after 24 h (For measured values see table below)

Details on test conditions:

TEST MEDIUM / WATER PARAMETERS:
- Source: Tap water
- Holding water different from dilution water: No

TEST SYSTEM:
- Renewal of test solution: daily
- Volume of test vessels: 3 L
- Volume/animal: 0.43 l/fish
- Number of animals/vessel: 7
- Number of vessels/ concentration: 1
- Test performed in closed vessels due to significant volatility of TS: No

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: Light/dark cycle 16 h / 8 h

SAMPLING
- Mortality recorded daily
- pH, temperature and O2 concentration checked at the beginning of incubation and daily after 24 h

STATISTICS
- The LC0 and LC100 were directly taken from the concentrations being tested. LC50 determined as geometric mean between LC0 and LC100.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

50 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

71 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- No other effects were observed in treatment groups and no dead animals were observed in the non-tretaed control group

Any other information on results incl. tables

Sublethal observations / clinical signs:

- Mortality data of exposed animals:

Test substance Concentration (nominal/) [mg/l]	Mortality
	Number				Percentage
	24 h	48 h	72 h	96 h	24 h	48 h	72 h	96 h
control	No data				0	0	0	0
100					57	100	100	100
50					0	0	0	0
25					0	0	0	0
12.5					0	0	0	0
6.25					0	0	0	0
Temp. [°C]	20.0-21.5	21.0-21.1	22.2-22.9	22.6
pH	7.61-8.40	7.52-8.44	7.76-8.29	8.03-8.14
Oxygen [mg/l]	7.7-7.9	7.2-8.1	7.7-8.1	7.8-8.1

Applicant's summary and conclusion

Conclusions:

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

Executive summary:

The substance was found to be acutely toxic towards Brachydanio rerio. A steep concentration-response relationship can be established from the study results. Exposure at 100 mg/L resulted in 100% mortality within 48 hours. At 50 mg/L no mortality was observed until test termination. The LC50 was calculated as the geometric mean between LC0 and LC100.

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. Possible substance losses due to biodegradation are unlikely in the semistatic system.

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 ≤10%. Therefore, the nominal concentrations can be considered as effective concentrations.

The temperature range over the test period (range 20.0 – 22.9 °C) was slightly out of the range (within 2°C) requested by the Guideline. This small deviation is not considered to influence the integrity of the test results.