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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report date:
1996

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
DIN 38412-8 (Pseudomonas Zellvermehrungshemmtest)
Deviations:
no
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
Reaction mass of 1-(2,3-epoxypropoxy)-2,2-bis ((2,3-epoxypropoxy)methyl) butane and 1-(2,3-epoxypropoxy)-2-((2,3-epoxypropoxy)methyl)-2-hydroxymethyl butane
EC Number:
701-135-4
Cas Number:
30499-70-8
Molecular formula:
C15H26O6;C15H22O5
IUPAC Name:
Reaction mass of 1-(2,3-epoxypropoxy)-2,2-bis ((2,3-epoxypropoxy)methyl) butane and 1-(2,3-epoxypropoxy)-2-((2,3-epoxypropoxy)methyl)-2-hydroxymethyl butane
Test material form:
other: liquid

Sampling and analysis

Analytical monitoring:
not specified

Test solutions

Vehicle:
no
Details on test solutions:
A stock solution (emulsion) of 12,500 mg per litre in Milli-Q water (tap water purified by reverse osmosis and subsequently passed over activated
carbon and ion-exchange cartridges (Millipore Corp., Bedford, Mass., USA)) was prepared. Ultra sonication and thorough mixing were performed to maximise the dissolving of the test substance. No additives were used.
The pH of the stock solution was 7.9.
A dilution series of the test substance stock solution was prepared.

Test organisms

Test organisms (species):
Pseudomonas putida
Details on inoculum:
Pseudomonas putida. Strain MIGULA, Berlin 33/2 (DSM 50026).
Source: BGA, Berlin, Germany.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
18 h

Test conditions

Test temperature:
23 +/- 1°C
Nominal and measured concentrations:
313 - 10,000 mg/l (nominal)
Details on test conditions:
EXPERIMENTAL CONDITIONS AND SOLUTIONS
Vessels: 100 ml Erlenmeyer flasks
Incubation temperature: 23 +/- 1°C
Incubation time:
- preliminary suspension cultures: 6 +/- 1 hours
- exposure: 18 +/- 2 hours

Stock solution I:
10.0 g NaN03
2.4 g K2HP04
1.2 g KH2P04
1.0 g yeast extract
Dissolved in 500 ml Milli-Q water.

Stock solution II:
10.0 g NaN03
2.4 g K2HP04
1.2 g KH2P04
Dissolved in 500 ml Milli-Q water.

Stock solution III:
40.0 g D(+)- glucose
Dissolved in 500 ml Milli-Q water.

Stock solution IV:
0.01 g Fe (III) citrate
4.0 g MgSD4 . 7H20
Dissolved in 1 1 Milli-Q water.

Culture medium (stock)
Nutrient medium: 18 g of agar dissolved in water, plus 50 ml of solution I, 125 ml of solution III and 100 ml of solution IV, made up to one litre with water.
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol

Results and discussion

Effect concentrationsopen allclose all
Duration:
18 h
Dose descriptor:
EC10
Effect conc.:
6 310 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Duration:
18 h
Dose descriptor:
EC50
Effect conc.:
> 10 000 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Results with reference substance (positive control):
The EC50 value of 3,5-dichlorophenol determined for Pseudomonas putida as calculated by linear regression (Y = 3.8772 X -62.5430) was 29 mg/l.
Reported statistics and error estimates:
After incubation extinction values were measured and the cell multiplication inhibition for each test concentration was determined.
When the concentration of the test substance was correlated with the corresponding inhibition by linear regression analysis, the following
equation was obtained:
Y = 14.4333 x - 44.7633
Where x represents the log concentration of TKK 30174, and Y the % inhibition.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
Based on the results of the test presently performed, the toxicity threshold value (EC10) of TKK 30174 was 6310 mg/l.
The EC50 was over 10,000 mg/l.
Executive summary:

TKK 30174, ARALDITE DY 0396 was investigated for its ability to inhibit the cell multiplication of the bacteria species Pseudomonas putida according to DIN 38412 Part L 8 (March 1991) and NEN-EN-ISO 10712 (1996). Suspensions of Pseudomonas putida were exposed to concentrations ranging from 312.5 to 10,000 mg TKK 30174  per litre. After incubation the extinction values of the bacterial cell suspensions were measured. Based on the results of the test presently performed, the toxicity threshold value (EC10) of TKK 30174 was 6310 mg/1. The EC50 was over 10,000 mg/l.


The EC50 value for the reference substance, 3,5-dichlorophenol, tested during the same experiment was 29 mg/1. Since this was within the accepted range of 10-30 mg/1 for the EC50 and the multiplication factor of the bacteria within the test was over 60, it was concluded that the test conditions were optimal and the results obtained were valid.