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Ecotoxicological information

Toxicity to microorganisms

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Reference
Endpoint:
toxicity to microorganisms
Type of information:
migrated information: read-across based on grouping of substances (category approach)
Adequacy of study:
key study
Study period:
not reported
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: National guideline study with acceptable restrictions
Qualifier:
according to guideline
Guideline:
other: Bewertung wassergefährdender Stoffe, III Bestimmung der akuten Bakterientoxizität
Qualifier:
equivalent or similar to guideline
Guideline:
other: ISO 10712
GLP compliance:
yes
Analytical monitoring:
yes
Details on sampling:
- Sampling method: 50 mL of the test substance stock solutions were taken after filtration of the saturated stock solution
- Sample storage conditions before analysis: frozen at -20 °C
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A saturated test substance solution with a nominal concentration of 10 mg/L was prepared by sonication for 30 minutes and a subsequent 24 h lasting mixing with a magnetic stirrer. Afterwards the solution was filtered and neutralised to a pH of 7.1 with NaOH
- Controls: 10 with inoculum without test substance to determine turbidity and for each test concentration 1 control without inoculum to determine possibly occuring colouration
Test organisms (species):
Pseudomonas putida
Details on inoculum:
- Preparation of inoculum for exposure: Bacteria from 7 days old stock culture were inoculated in a fluid nutrient medium in Erlenmeyer flasks and incubated for 16-20 h at 25 °C. Afterwards the extinction of the monochromatic radiation at 436 nm was determined for a 10 mm layer of the bacterial suspension by photoelectric measurement.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
16 h
Remarks on exposure duration:
16 - 20 h
Test temperature:
23 - 27 °C
pH:
pH was neutralised to 7.1 with NaOH
Nominal and measured concentrations:
nominal: 3.9, 7.8, 15.6, 31.3, 62.5, 125.0, 250.0, 500.0, 1000.0, 2000.0, 4000.0, 8000.0 mg/L (mean measured)
actual: (0.11 % of the nominal concentrations)
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): closed with aluminum foil caps
- Material, size, headspace, fill volume: 300 mL Erlenmeyer flasks
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Milli-Q water: water was first purified by reverse osmosis and thereafter passed over activated carbon and ion exchange cartridges

OTHER TEST CONDITIONS
- Adjustment of pH: with NaOH to 7.1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : extinction of monochromatic radiation at 436 nm in a 10 mm layer after 16 - 20 h
Reference substance (positive control):
yes
Remarks:
Methanol
Duration:
18 h
Dose descriptor:
EC10
Effect conc.:
883 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Remarks on result:
other: given as TT (toxicity threshold)
Results with reference substance (positive control):
- Other: The toxicity threshold value of the reference substance was in the usual range known at RCC Notox

Table 1: Extinction of the test substances solutions after 16 - 20 h

Concentration

[mg/L]

Extinction at 436 nm

Replicate I

Replicate II

Replicate III

Arithm. mean of replicates

Replicate without inoculum

Corrected arithmetic mean*

 

3.9

0.822

0.910

0.927

0.886

0.001

0.885

 

7.8

0.760

0.745

0.827

0.777

0.000

0.777

 

15.6

0.801

0.836

0.842

0.826

0.000

0.826

 

31.3

0.823

0.829

0.843

0.832

0.000

0.832

 

62.5

0.849

0.815

0.827

0.830

0.000

0.830

 

125.0

0.845

0.816

0.832

0.831

0.000

0.831

 

250.0

0.843

0.834

0.833

0.837

0.000

0.837

 

500.0

0.796

0.807

0.852

0.818

0.000

0.818

 

1000.0

0.775

0.740

0.699

0.738

0.001

0.737

 

2000.0

0.672

0.658

0.654

0.661

0.009

0.652

 

4000.0

0.680

0.694

0.703

0.692

0.024

0.668

 

8000.0

0.746

0.731

0.781

0.753

0.057

0.696

 

* corrected with extinction values from replicate without inoculum

 

The resulting nominal toxicity threshold (TT), determined graphically at 883 mg prod./L. However, due to the DOC and TCC measurements only 0.11 % of the added test substance persist in the test medium. Thus, the actual TT (which is equivalent to the EC10) must be 0.97 mg prod./L.

Table 2: Extinction after 16 - 20 h in the reference substance and the blank control

Concentration

[mg prod./L]

Extinction at 436 nm

Reference substance

Blank controls

3950.0

0.775

0.855

7900.0

0.695

0.901

15800.0

0.490

0.912

31600.0

0.220

0.903

63200.0

0.057

0.830

 

0.810

0.790

0.788

0.803

0.809

Arithmetic mean

0.840

With respect to the historical test with the reference substance methanol, it can be concluded that the test conditions were optimal and the results are valid.

Description of key information

No toxic effects on microorganisms were observed up to the limit of water solubililty, no PNEC is derived. 

Key value for chemical safety assessment

Additional information

Within the fatty acid category studies describing the toxicity of fatty acids to Pseudomonas putida are available. For the mixture under discussion read-across is performed to a study conducted with stearic acid C18 (CAS 57 -11 -4). The cell multiplication inhibition test was conducted following GLP-requirements. The used method in the test was similar to the ISO guideline 10712. In the study an EC10 (18 h) of 883 mg/L (nominal) (Coenen, 1989) was reported. Concluding from the results of this study the considered mixture is regarded to be not toxic to microorganisms. As no toxic effects on microorganisms were observed up to the limit of water solubililty, no PNEC is derived.