1,4:3,6-dianhydro-2,5-di-O-methyl-D-glucitol

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:

05 Dec 2016 - 11 Apr 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Medicines & Healthcare products Regulatory Agency, UK

Test material

Test material information

Constituent 1

Reference substance name:

1,4:3,6-dianhydro-2,5-di-O-methyl-D-glucitol

EC Number:

226-159-8

EC Name:

1,4:3,6-dianhydro-2,5-di-O-methyl-D-glucitol

Cas Number:

5306-85-4

Molecular formula:

C8H14O4

IUPAC Name:

1,4:3,6-dianhydro-2,5-di-O-methyl-D-glucitol

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: control and 100 mg/L
- Sampling method: duplicate samples were taken at 0 and 72 hours of the test.
- Sample storage conditions before analysis: The samples were stored frozen prior to analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A 100 mg/L stock solution of the test substance was prepared by dissolving 100 mg of test substance in 1 L of dilution water.
- Eluate: no
- Differential loading: no
- Controls: yes, test medium control

Test organisms

Test organisms (species):

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

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Strain: CCAP 278/4
- Source (laboratory, culture collection): Culture Collection of Algae and Protozoa (CCAP), SAMS Research Services Ltd, Scottish Marine Institute, Oban, Argyll, Scotland.
- Method of cultivation: Master cultures were maintained in the laboratory by the periodic replenishment of culture medium. The master cultures were maintained under constant aeration and illumination at 21 ± 2 °C.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

72 h

Post exposure observation period:

none

Test conditions

Test temperature:

24 ± 1 ºC

pH:

Control:
7.6 at 0 hours
8.4 at 72 hours

Test concentration 100 mg/L:
7.7 at 0 hours
8.5 at 72 hours

Nominal and measured concentrations:

0 and 100 mg/L (nominal concentration)
0 and 98.7 mg/L (measured concentrations at T = 0 h); 0 and 97.8 mg/L (at T = 72 h)

Details on test conditions:

TEST SYSTEM
- Test vessel: 250 mL glass conical flasks
- Type (delete if not applicable): closed with polyurethane foam caps
- Material, size, headspace, fill volume: 250 mL
- Aeration: no, but constantly shaken at 150 rpm
- Initial cells density: 5 x 10^3 cells per mL
- Control end cells density: 8.2 x 10^5 cells per mL (mean)
- No. of vessels per concentration (replicates): 6
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reverse osmosis purified deionized water and the pH adjusted to 7.5 with NaOH or HCl.

OTHER TEST CONDITIONS
- Sterile test conditions: no
- Adjustment of pH: yes
- Photoperiod: continuous illumination
- Light intensity and quality: ca. 7000 lux; wave 380 – 730 nm

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: Cell density was measured at 24, 48 and 72 hour using Coulter® Multisizer Particle Counter.


TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 0.10, 1.0, 10, 100 mg/L
- Results used to determine the conditions for the definitive study: Maximum test concentration (100 mg/L) had no effect on algae growth.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 3

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 4

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 5

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 6

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

Growth rate and yield of green algae P. subcapitata were not affected by the test item at a nominal test concentration of 100 mg/L over the 72-hour exposure period.

Results with reference substance (positive control):

72 h ErC50 = 1.4 mg/L; NOECr = 0.25 mg/L
72 h EyC50 = 0.6 mg/L; NOECy = 0.25 mg/L

Reported statistics and error estimates:

Student’s t-test incorporating Bartlett's test for homogeneity of variance (Sokal and Rohlf, 1981).

Any other information on results incl. tables

Cell Densities and Percentage Inhibition of Growth from the Range-finding Test:

Nominal Concentration (mg/L)	Mean Cell Densities (cells per mL)		Inhibition Values (%)
	0 h	72 h	Growth rate	Yield
control	9.06E+03	1.46E+06	-	-
0.1	9.03E+03	1.25E+06	4	14
1	8.61E+03	1.34E+06	1	8
10	9.22E+03	1.26E+06	4	14
100	9.40E+03	1.27E+06	4	13