Registration Dossier

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

Constituent 1
Chemical structure
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:
(3R,3aR,6S,6aR)-3,6-dimethoxy-hexahydrofuro[3,2-b]furan

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 concentrationsopen allclose all
Duration:
6 h
Dose descriptor:
EC50
Effect conc.:
> 100 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
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.

Applicant's summary and conclusion