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

Toxicity to microorganisms

Administrative data

Endpoint:

toxicity to microorganisms, other

Remarks:

growth inhibition of Pseudomonas putida

Type of information:

experimental study

Adequacy of study:

key study

Study period:

3 Mar 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Cell multiplication inhibition test with P. putida with limitations in methodology (method Bringmann and Kühn 1980), exposure time 6 h.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Methodology followed Bringmann and Kuehn (1980): Comparison of the toxicity threshold of water pollutants to bacteria, algae and protozoa in the cell multiplication inhibition test, Water Research 14, 231-241, with modification described by Slabbert (1986): Improved bacterial growth test for rapid water toxicity screening, Bull. Environ. Contam. Toxicol. 37, 565-569.

GLP compliance:

yes

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

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION:
Stock solution of 500 mg/L of the test substance in water was prepared. Diluted solutions were used for testing.

Test organisms

Test organisms (species):

Pseudomonas putida

Details on inoculum:

- Source: national Collections of Industrial and Marine Bacteria Ltd, Aberdeen, UK
- Preparation of inoculum for exposure: the freeze-dried culture was rehydrated in 0.5 mL of nutrient broth. A loop of this suspension was streaked into a nutrient agar slope in a universal bottle. Incubation at 25 °C took place for 24 hours. The culture was stored at laboratory temperature until use.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

6 h

Post exposure observation period:

none

Test conditions

Hardness:

not stated

Test temperature:

25.0 +/- 0.5 °C

pH:

not stated

Dissolved oxygen:

not stated

Nominal and measured concentrations:

0 and 100 mg/L (nominal concentration)

Details on test conditions:

TEST SYSTEM
- Test vessel: sterile conical flasks, volume 50 mL, shaken at 150 rpm for 6 hours

No of replicates:
- No. of vessels per control (no test substance + inoculum): 3
- No. of vessels per positive control (replicates): 3
- No. of vessels per test concentration (test substance + inoculum): 3
- No vessels per blank control (no test substance, no inoculum): 1
- No. of vessels per abiotic control (test substance, no inoculum): 1
- Initial absorbance: 0.8 +/- 0.05 at 600 nm

TEST MEDIUM / WATER PARAMETERS
Basal salt solution - dissolved in 1 L of deionised water:
26.25 g dipotassium hydrogen phosphate
11.25 g potassium dihydrogen phosphate
1.175 g trisodium citrate dihydrate
2.5 g ammonium sulphate
0.25 g magnesium sulphate heptahydrate
Glucose solution - dissolved in 1 L of deionised water: 6.25 g glucose
Equal volumes of basal salt solution and glucose solutions were mixed together.

EFFECT PARAMETERS MEASURED: optical density at 600 nm (Uvikon 930 spectrophotometer)

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol (purity 99%)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

6 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Effect concentrations 2

Duration:

6 h

Dose descriptor:

EC10

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Details on results:

No growth inhibition was measured in any of the test solution. The EC10 and EC50 values for the test substance are above 100 mg/L (nominal).

Results with reference substance (positive control):

96% inhibition of growth was observed at 18 mg/L of reference substance.