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Toxicity to microorganisms

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Reference
Endpoint:
toxicity to microorganisms
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1992-04-02 to 1992-04-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Qualifier:
according to guideline
Guideline:
DIN 38412-8 (Pseudomonas Zellvermehrungshemmtest)
Deviations:
no
GLP compliance:
yes
Analytical monitoring:
no
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Test 1: 1.7 g of the test substance was dissolved in 1 L purified water to assess effects of test item without adjusted pH within range of maximum solubility (stock solution). Appropriate amounts of the stock solution were diluted (f:2) to give a series of 10 dilutions, the highest concentration of the test article corresponding to 850 mg/L. In Test 2, the same amount of test article was used after correction of the stock solution which was done to distinguish whether inhibition is caused by test article or by an extreme pH value produced by test article. The pH of the stock solution was mesured 1 h after preparation. Thereafter, the solution was adjusted to a pH of 6.85 with NaOH (1 mol/L) from which a series of nine dilutions (f:1.25, f:2) was prepared. The highest concentration was 1360 mg/L. The pH value of the solution did not remain constant. During the performance of the test, the pH value decreased rapidly.
Test organisms (species):
Pseudomonas putida
Details on inoculum:
- Laboratory culture: DSM-No 50026 (lyophilisate, March 1992)
- Preparation of inoculum for exposure: three 250 mL Erlenmeyer flasks with 100 mL nutritive solution for pre-cultures were each inoculated with stock culture maximally 7 days old. After incubating for 18 ± 2 h at 21°C, the turbidity of the bacterial suspension was measured and adjusted to FNU 100 by addition of medium for pre-culture. This suspension was used as inoculum for the pre-culture. A pre-culture with an initial turbidity of FNU 10 was prepared by inoculating 90 mL of medium for pre-culture with 10 mL of bacterial suspension (FNU 100). Afer incubating for 7 h at 21°C ± 1°C, the turbidity was remeasured and adjusted to FNU 50 by addition of test medium. This suspension was used as inoculum for the test culture.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
17 h
Test temperature:
21 ± 1 °C
pH:
Test 1: 2.84
Test 2: 6.85 (immediately after pH adjustment)
Nominal and measured concentrations:
Nominal:
test 1: 1.7, 3.3, 6.6, 13, 27, 53, 106, 213, 425, 850 mg/L
test 2: 6.6, 13, 27, 53, 106, 213, 425, 850, 1360 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: Erlenmeyer flasks
- Material, size, headspace, fill volume: 250 mL (size)
- Aeration: none during the test
- No. of vessels per concentration: 3
- No. of vessels per control: 3


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: purified water
- Nutrient solution: in accordance to the guideline

OTHER TEST CONDITIONS
- Adjustment of pH: test 1 no adjustment of ph; test 2: adjustment to pH 6.85

EFFECT PARAMETERS MEASURED: cell multiplication

TEST CONCENTRATIONS
- Spacing factor for test concentrations: dilution of f.2 (test t) and f.1,5 and f:2 (test 2)
Reference substance (positive control):
yes
Remarks:
3,5-dichlorphenol
Duration:
17 h
Dose descriptor:
EC10
Effect conc.:
ca. 26 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Remarks on result:
other: without adjusted pH (pH 2.84)
Duration:
17 h
Dose descriptor:
EC50
Effect conc.:
ca. 80 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Remarks on result:
other: without adjusted pH (pH 2.84)
Duration:
17 h
Dose descriptor:
EC10
Effect conc.:
ca. 44 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Remarks on result:
other: adjusted pH (pH 6.85)
Duration:
17 h
Dose descriptor:
EC50
Effect conc.:
ca. 90 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth inhibition
Remarks on result:
other: adjusted pH (pH 6.85)
Details on results:
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none reported
- Effect concentrations exceeding solubility of substance in test medium: none reported
Results with reference substance (positive control):
- Results with reference substance valid? yes
- Relevant effect levels: The reference item exerted a pronounced inhibitory effect on cell growth which resulted in EC50 and EC10 values of 20.6 and 5.8 mg/L, respectively.
Reported statistics and error estimates:
Inhibition of cell multiplication is plotted as a function of test article concentration (fitted with spline) and the effective concentrations EC10 and EC50 are calculated from the graphic representation.

Description of key information

The determined EC10 was 26 mg/L (without pH adjustment)

Key value for chemical safety assessment

EC50 for microorganisms:
26 mg/L

Additional information

The toxicity of 1,3-propanesultone to microorganisms was tested according to German Standard guideline DIN 38412 L8. In a 17-hr acute toxicity test suspensions of Pseudomonas putida were exposed to the test item at maximum concentrations of 850 mg/L (test without adjustment of pH) and 1360 mg/L (test with adjustment of pH). The determined EC10 was 26 mg/L (test without adjustment of pH).