1-methyltrimethylene dimethacrylate

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020-09-03 - 2020-09-18

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

Guideline study OECD 201, GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

2011

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Version / remarks:

No. 266/2016

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

TEST MATERIAL:
- Name of test material: 1,3-BDDMA
- CAS No. 1189-08-8
- EC No. 214-711-0

Analytical monitoring:

yes

Details on sampling:

All concentration levels and the control were analytically verified via GC-MS at the start (0 hours) and at the end of the exposure (72 hours) with algae. At the start, samples were taken from one additional replicate of each test item concentration and the control and at the end samples were taken from pooled replicates.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
stock solution: 100 mg/L, freshly prepared with dilution water
The dispersion was treated with ultrasound for 5 minutes and stirred for 30 minutes (1100 rpm, room temperature) with a magnetic stirrer. The stock solution was checked via laser beam (Tyndall effect) for undissolved test item. The Tyndall effect was negative.
Out of the stock solution five concentrations were prepared in a geometric series with a dilution factor of 10, i.e. 1.00, 3.16, 10.0, 31.6, 100 mg test item/L.

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata
- Strain: CCAP 278/4
- Source (laboratory, culture collection): Culture Collection of Algae and Protozoa (CCAP),
AMS Research Services Ltd
Dunstaffnage Marine Laboratory
Dunbeg, OBAN; Argyll PA37 1QA; Scotland, UK
- Age of inoculum (at test initiation): The four-days old preculture, prepared in dilution water was used as inoculum
- Method of cultivation: Fresh stocks prepared every month on Z-Agar; Light intensity amounts 2567 – 5130 lux for 24 hours per day.

ACCLIMATION
- Acclimation period: no data
- Culturing media and conditions (same as test or not): according to the guideline (OECD 201 medium)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

21-24 °C

pH:

initial: 7.75 to 7.88
final: 8.20 to 8.96

Nominal and measured concentrations:

nominal: 1.00, 3.16, 10.0, 31.6, 100 mg/L
measured: 0.988, 3.02, 9.90, 31.5, 96.3 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: Sterile Erlenmeyer flasks, volume: 250 mL, sealed with cotton wool plugs, test volume 100 mL
- Aeration: agitation, Test containers were placed on a rotary shaker and oscillated at approximately 70 rpm
- Initial cells density: 5375 cells/mL
- Control end cells density: mean 607626 cells/mL
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes, OECD 201 medium

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Photoperiod: 24 hours/day light
- Light intensity and quality: mean 6201 lux (CV 7.42%)

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Cell density was measured daily via Chlorophyll-a- fluorescence,excitation at 436 nm, emission at 685 nm

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 3.16
- Range finding study: 1.00, 10.00, 100.00 mg/l
- Test concentrations: 1.00, 3.16, 10.00, 31.60, 100.00 mg/L
- Results used to determine the conditions for the definitive study:
Growth inhibition 100% at 100.00 mg/L, 27% at 10.00 mg/L, -1% at 1.00 mg/L
Yield inhibition 100% at 100.00 mg/L, 72% at 10.0 mg/L, -4% at 1.00 mg/L

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

9.19 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

3.16 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

3.16 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

2.35 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

7.27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Results with reference substance (positive control):

- Results with reference substance are valid:
- ErC50: 1.12, 95 % confidence interval: 1.08 – 1.15 / valid range: 0.771 ± 0.563
- EyC50: 0.594, 95 % confidence interval: 0.518 – 0.664 / valid range: 0.420 ± 0.344

Reported statistics and error estimates:

EC10-, EC20- and EC50-values of the growth rate and yield inhibition after 72 hours were calculated by sigmoidal dose-response regression. Calculation of the confidence intervals EC10-, EC20- and EC50-values were carried out using standard procedures.

Mean cell densities [cells/mL] during the final test:

Nominal test item concentration[mg/L]	Exposure time [h]
	0	24	48	72
100	5375	n.a.	n.a.	n.a.
31.6	5375	5327	n.a.	n.a.
10.0	5375	10116	48667	287283
3.16	5375	12296	79617	460209
1.00	5375	16017	102872	685118
Control	5375	16502	105155	607626

  

Growth rate, yield during the final test: 

Nominal test item concentration [mg/L]	Growth rate [/d]	Rate-related inhibition [%]	Yield [cells/mL]	Inhibition of yield [%]
100	n.a.	100	n.a.	100
31.6	n.a.	100	n.a.	100
10.0	1.32	16	281908	53
3.16	1.48	6	9454834	24
1.00	1.61	-3	679743	-13
control	1.57		602251

Validity criteria fulfilled:

yes

Conclusions:

In this study 1,3-BDDMA was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours.
The NOEC, EC10 and EC50 values based on growth rate were 1.00 mg/L, 9.19 (95% c.i. 6.10 – 9.95) mg/L and 13.0 (95% c.i. 10.7 – 22.3) mg/L, respectively. 
The NOEC, EC10 and EC50 values based on yield were 1.00 mg/L, 2.35 (95% c.i. 1.56 – 3.52) mg/L and 7.27 (95% c.i. 5.20 – 10.2) mg/L, respectively.
The effect values given are based on the nominal concentrations of the test item.

Executive summary:

In a 72 hour acute toxicity study, the cultures of Pseudokirchneriella subcapitata were exposed to 1,3-Butanediol dimethacrylate (1,3 -BDDMA) at nominal concentrations of 0 (control), 1.00, 3.16, 10.00,31.60, 100.00 mg/L under static conditions in accordance with the OECD guideline 201 (Version 2011). 1,3-BDDMA was found to inhibit the growth of the freshwater green alga Pseudokirchneriella subcapitata after 72 hours with the following effect values (based on nominal test item concentrations):

Results Synopsis

Test Organism: Pseudokirchneriella subcapitata HINDÁK CCAP 278/4 (axenic)

Test Type: Static

 

72 hr ErC50: 13 mg/L              95% C.I.: 10.7 – 22.3 mg/L

72 hr EyC50: 7.27 mg/L             95% C.I.: 5.20 – 10.2 mg/L

 

72 hr ErC10: 9.19 mg/L              95% C.I.:  6.10 – 9.95 mg/L

72 hr EyC10:  2.35 mg/L             95% C.I.:  1.56 – 3.52 mg/L

 

72 hr NOEC: 1.00 mg/L              

 

Endpoint(s) Effected:  growth rate, yield

Description of key information

The 72 h EC10 of 1,3-BDDMA in freshwater algae was 9.19 mg/L (based on growth rate) (HMRTF, 2021)

The 72 h EC50 of 1.3-BDDMA in freshwater algae was 13.0 mg/L (based on growth rate) (HMRTF, 2021)

Key value for chemical safety assessment

EC50 for freshwater algae:

13 mg/L

EC10 or NOEC for freshwater algae:

9.19 mg/L

Additional information

One study in freshwater algae is available for 1,3-BDDMA, which is considered reliable without restriction (Klimisch score 1, guideline study, GLP).

In a 72 hour acute toxicity study, the cultures of Pseudokirchneriella subcapitata were exposed to 1,3-BDDMA at measured concentrations of 0 (control), 0.988, 3.02, 9.90, 31.5 and 96.3 mg/L under static conditions in accordance with the OECD guideline 201 (adopted 2011). 

Microscopic evaluation of the cells at start and end of the incubation period revealed no morphological abnormalities.

The NOEC, EC10 and EC50 values based on growth rate were 1.0 mg/L, 9.19 mg/L and 13.0 mg/L, respectively.