Diallyl 2,2'-oxydiethyl dicarbonate

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1983-04-04 to 1983-04-08

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

other: U. S. Environmental Protection Agency (1978): The Selenastrum capricornutum Printz Algal Assay Bottle Test

Deviations:

not specified

Principles of method if other than guideline:

Test was performed according to: U. S. Environmental Protection Agency (1978): The Selenastrum capricornutum Printz Algal Assay Bottle Test, Environmental Research Laboratory, Corvallis, OR. 126 pp.

GLP compliance:

yes

Analytical monitoring:

no

Remarks:

But due to good water solubility and slow hydrolysis the test item is expected to be stable for the duration of the test.

Details on sampling:

No analytical verification of test substance concentrations

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A primary stock solution of CR-39 was prepared by adding 0.87 mL of CR-39 to a 100mL volumetric flask and bringing to volume with triethylene glycol (TEG). Additional stocks were prepared by serial dilution of the primary stock solution and 0.05 mL of the appropriate stock added to each test flask.
- Controls: The solvent control received the equivalent volume of TEG as added to all test flasks (0.05 mL; 1000 ppm TEG concentration), but no test material. The growth medium control did not receive TEG or test material. An additional set of flasks containing 10000 ppm TEG was prepared and cell growth monitored simultaneously with the CR-39 definitive test. This treatment was conducted to show the deleterious effects of TEG on cell growth if greater volumes of TEG had been used in the test flasks to obtain higher CR-39 test concentrations.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): triethylene glycol (TEG).

Test organisms (species):

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

Details on test organisms:

TEST ORGANISM
- Common name: freshwater alga Selenastrum capricornutum
- Source (laboratory, culture collection): University of Texas, Austin, Texas,


ACCLIMATION
- Acclimation period: no data
- Culturing media and conditions (same as test or not): no data
- Any deformed or abnormal cells observed: no data

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

Not reported

Test temperature:

24 +/- 1 °C

pH:

Initial: 7.4
Final (after 96 h): 8.8 to 9.7.

Dissolved oxygen:

Not reported

Salinity:

Not reported

Nominal and measured concentrations:

Nominal concentrations: 0, 0.625, 1.25, 2.5, 5.0 and 10.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: Test flasks according to guideline
- Renewal rate of test solution (frequency/flow rate): No renewal
- Initial cells density: Beginning cell numbers in the test flasks were approximately 2.0 x 10E4 cells/mL
- Control end cells density: After 96 hours of exposure to CR-39, decreases of In vivo chlorophyll a were 12% and 6% in 0.625 ppm and 5.0 ppm, respectively. In vivo chlorophyll a increased 9, 35, and 6% in cultures exposed to 1.25, 2.5, and 10 ppm, respectively. Decreases in cell numbers occurred in all test concentrations and ranged from 5% in cultures exposed to 2.5 ppm to 10% in cultures exposed to 0.625 ppm
- No. of vessels per concentration (replicates): Three replicates were employed for each of the test concentrations
- No. of vessels per control (replicates):Three replicates were employed for the controls.
- No. of vessels per vehicle control (replicates): 3 replications with 10000 µL/L triethylene glycol (TEG) [1000 ppm TEG]

GROWTH MEDIUM
- Standard medium used: yes; test medium was Algal Assay Procedure medium.

OTHER TEST CONDITIONS
- Sterile test conditions: No information available
- Adjustment of pH: at test initiation pH 7.8
- Photoperiod: constant illumination
- Light intensity and quality: 4500 lux, constant illumination

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Change in cell number;
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]: Cell counts were made by using a hemacytometer and a Zeiss Standard 14 compound microscope.
- Chlorophyll measurement: Measurements of in-vivo chlorophyll were made using a Turner Model 111 fluorometer.

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Not relevant
- Justification for using less concentrations than requested by guideline: Not relevant
- Range finding study
- Test concentrations: No range finding study

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

cell number

Details on results:

- Observation of abnormalities (for algal test): None reported
- Any stimulation of growth found in any treatment: In vivo chlorophyll-a increased 9, 35, and 6% in cultures exposed to 1.25, 2.5, and 10 ppm, respectively

Results with reference substance (positive control):

No reference substance used

Reported statistics and error estimates:

No statistical procedures reported.

Table 1	In vivo chlorophyll α, (expressed in relative fluorescence units determined with a Turner Model 111 fluorometer) during a 4-day continuous exposure of Selenastrum capricornutum to CR-39. Values are means of three flasks. Standard deviations (±1 S.D.) are in parentheses. Percentage change is increase or decrease of relative fluorescence units in exposed cultures as compared to the solvent control at day 4.
Nominal concentration (mg/l; ppm)		Relative fluorescence units				Percentage change
		Day 1	Day 2	Day 3	Day 4	Day 4
Growth medium control		23 (1)	91 (7)	263 (61)	767 (81)	-7
Solvent control		22 (2)	132 (19)	373 (47)	827 (76)	---
0.625		21 (2)	84 (10)	243 (25)	727 (42)	-12
1.25		22 (1)	122 (17)	390 (46)	900 (35)	9
2.5		20 (2)	125 (24)	457 (75)	1,120 (178)	+35
5.0		23 (0)	100 (3)	353 (15)	780 (72)	-6
10		22 (1)	110 (12)	403 (45)	880 (92)	+6
10,000 µl TEG/l		21 (1)	134 (2)	80 (20)	520 (69)	-37

Table 2	Results of a 96-hour exposure of Selenastrum capricornutum to CR-39. Cells per milliliter are the means from triplicate flasks after 96 hours of exposure. Percentage change is decrease of cell numbers in exposed cultures as compared to the solvent control after 96 hours of exposure.Standard deviations (±1 S.D.) are in parentheses.
Nominal concentration (mg/l; ppm)		Cells/ml (x 10^4)	Percentage change	pH
				Initial	Final
Growth medium control		217 (12)	-14	7.4	9.7
Solvent control		253 (16)	----	7.4	9.6
0.625		227 (25)	-10	7.4	9.5
1.25		237 (23)	-6	7.4	9.5
2.5		241 (26)	-5	7.4	8.9
5.0		231 (16)	-9	7.4	9.6
10.0		229 (12)	-9	7.4	9.4
10,000 µl TEG/l		141 (23)	-44	7.4	8.8

 

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (96 hours) was >10 mg/L.

Executive summary:

A phytotoxicity test was conducted at EG&G Bionomics Marine Research Laboratory (BMRL), Pensacola, Florida, to determine the effect of CR-39® on the freshwater alga Selenastrum capricornutum. Test was performed according to: U. S. Environmental Protection Agency (1978): The Selenastrum capricornutum Printz Algal Assay Bottle Test, Environmental Research Laboratory, Corvallis, OR. 126 pp. The EC50 (96 hours) was >10 mg/L.

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982-12-22 to 1982-12-26

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: Bioassay procedures for the ocean disposal permit program; EPA-600/9-78-010. 121 pp

Deviations:

not specified

Principles of method if other than guideline:

Test was performed according to: U.S. Environmental Protection Agency. 1978. Bioassay procedures for the ocean disposal permit program. EPA-600/9-78-010. 121 pp.

GLP compliance:

yes

Analytical monitoring:

no

Remarks:

But due to good water solubility and slow hydrolysis the test item is expected to be stable for the duration of the test.

Details on sampling:

No analytical verification of test substance concentrations

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A primary stock solution of CR-39 was prepared by adding 0.87 mL of CR-39 to a 100mL volumetric flask and bringing to volume with triethylene glycol (TEG).
- Controls: The solvent control received the equivalent volume of TEG as added to all test flasks (0.05 mL; 1000 ppm TEG concentration), but no test material.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): triethylene glycol (TEG).

Test organisms (species):

Skeletonema costatum

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

None

Hardness:

Not reported

Test temperature:

20 °C

pH:

Initial: 8.1
Final (after 96 h): 8.7 to 8.9

Dissolved oxygen:

Not reported

Salinity:

Artificial seawater (Rila Marine Mix) adjusted to approximately 30 parts per thousand salinity and enriched with nutrients.

Nominal and measured concentrations:

Nominal concentrations: 0, 0.312, 0.625, 1.25, 2.5, 5.0 and 10.0 µL/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: Test flasks according to guideline
- Renewal rate of test solution (frequency/flow rate): No renewal
- Initial cells density: Beginning cell numbers in the test flasks were approximately 2.0 x 10e4 cells/mL
- No. of vessels per concentration (replicates): Three replicates were employed for each of the test concentrations
- No. of vessels per control (replicates):Three replicates were employed for the controls.
- No. of vessels per vehicle control (replicates): 3 replications with 1000 mg/L triethylene glycol (TEG)


GROWTH MEDIUM
- Standard medium used: yes; test medium was Algal Assay Procedure medium.


OTHER TEST CONDITIONS
- Sterile test conditions: No information available
- Adjustment of pH: at test initiation pH 8.1
- Photoperiod: constant illumination
- Light intensity and quality: 4300 lux, constant illumination



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Change in cell number;
- Determination of cell concentrations: [counting chamber; electronic particle counter; fluorimeter; spectrophotometer; colorimeter]: Cell counts were made by using a hemacytometer and a Zeiss Standard 14 compound microscope.
- Chlorophyll measurement: Measurements of in-vivo chlorophyll were made using a Turner Model 111 fluorometer.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: Not relevant
- Justification for using less concentrations than requested by guideline: Not relevant
- Range finding study
- Test concentrations: No range finding study

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 10 other: µL/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

cell number

Remarks on result:

other: = 11.48 mg/L

Results with reference substance (positive control):

No reference substance used

Reported statistics and error estimates:

No statistical procedures reported.

Validity criteria fulfilled:

yes

Conclusions:

The EC50 (96 hours) was >10 µL/L [= 11.48 mg/L]

Executive summary:

A phytotoxicity test was conducted at EG&G Bionomics Marine Research Laboratory (BMRL), Pensacola, Florida, to determine the effect of CR-39 on the marine alga Skeletonema costatum. Test was performed according to U.S. Environmental Protection Agency (1978): Bioassay procedures for the ocean disposal permit program. EPA-600/9 -78 -010 (121 pp). Test medium was artificial seawater (Rila Marine Mix) adjusted to approximately 30 parts per thousand salinity and enriched with nutrients. The obtained EC50 (96 hours) was >10 µL/L [ = 11.48 mg/L].

Description of key information

- EC50 (96h) > 10 mg/L for Selenastrum capricornutum (guideline equivalent or similar to EPA OPP 72-2 (Selenastrum capricornutum Printz Algal Assay Bottle Test) EC50 (96h) > 11.48 mg/L for marine water algae [test followed internal procedures of the laboratory (Static acute Mysid shrimps toxicity test, June 1982)

- EC50 (96h) > 10 µg/L (=11.48 mg/L) for Skeletonema costatum (test equivalent or similar to Bioassay procedures for the ocean disposal permit program, EPA-600/9 -78 -010, 121)

Key value for chemical safety assessment

EC50 for freshwater algae:

10 mg/L

EC50 for marine water algae:

11.48 mg/L

Additional information

This endpoint is filled from data from one study in Selenastrum capricornutum (freshwater algae) (Maziarz, 1983a) and another in Skeletonema costatum (saltwater algae) (Maziarz,1983b). The no observable effect concentration (NOEC) for 'diallyl 2,2'-oxydiethyl dicarbonate' in Selenastrum capricornutum is 10 mg/L and in Skeletonema costatum 10µL/L (=11.48 mg/L).