2,2,4(or 2,4,4)-trimethylhexane-1,6-diamine

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to microorganisms, other

Remarks:

Pseudomonas putida

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993-02-23 to 1993-02-26

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

other: DIN 38412, part 8

Deviations:

not specified

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION
- Stock solution: 6.25 g/l in aqua bidest.
- Nutrient solutions: According to guideline, no details reported
- Bacteria suspension: 436 nm extinction 0.156-0.160, adjusted to 0.150  with culture medium
- Addition of  80 ml stock solution (diluted at six levels) or aqua bidest., 2.5 ml nutrient solution II,  2.5 ml nutrient solution III, 5 ml nutrient solution 
IV, 10 ml bacteria suspension or aqua bidest.

Test organisms (species):

Pseudomonas putida

Details on inoculum:

TEST ORGANISMS
- Strain: Pseudomonas putida MIGULA

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

16 h

Remarks on exposure duration:

+/- 1 hour

Test temperature:

20.7-21.3 °C

pH:

7.2 (25 mg/l) - 8.6 (200 mg/l); not adjusted

Nominal and measured concentrations:

25; 50; 75; 100; 150; 200 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Exposure vessel type: Sterile 250 ml Erlenmeyer flasks with cellulose  plugs
- Number of replicates: 2 independent tests with each 3 replicates per exposure concentration, 5 growth controls (no test substance), 6 sterile 
controls (no test organisms)

TEST PARAMETER
- light absorption at 436 nm, determined with Zeiss  photometer PM 6: test solutions vs. sterile controls; growth controls vs. deionized water

STATISTICAL METHODS: Multiplan computer program "MIGULA" (regression analysis)

Reference substance (positive control):

not specified

Duration:

17 h

Dose descriptor:

EC10

Effect conc.:

72 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: mean value

Duration:

17 h

Dose descriptor:

EC50

Effect conc.:

89 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: mean value

Details on results:

Test 1: EC10 = 76.1 mg/l, EC50 = 92.0 mg/l
Test 2: EC10 = 67.5 mg/l, EC50 = 85.9 mg/l

Validity criteria fulfilled:

not specified

Conclusions:

The EC50 (16 hours) was determined as 89 mg/L indicating that the test substance may be harmful to aquatic microorganisms.

Executive summary:

The effects of the test item on the bacteria species Pseudomonas putida were investigated by Hüls AG (1993) according to DIN 38412, part 8 (national German guideline). Six concentrations + control were tested over a period of 16 hours.

The EC50 was determined as 89 mg/L and the EC10 as 72 mg/L.

The study was assessed as "reliable without restrictions".

Reference 2

Endpoint:

toxicity to microorganisms

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1992-12-01 to 1992-12-04

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

DIN 38412-8 (Pseudomonas Zellvermehrungshemmtest)

Principles of method if other than guideline:

Method: other: DIN 38412 part 8

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION
- Stock solution: 12.5 and 8.75 g/l in aqua bidest.
- Nutrient solutions: According to guideline, no details reported
- Bacteria suspension: 436 nm extinction 0.152-0.155, adjusted to 0.150  with culture medium
- Addition of  80 ml stock solution (diluted at six levels) or aqua bidest. 2.5 ml nutrient solution II;  2.5 ml nutrient solution III; 5 ml nutrient solution IV   10 ml bacteria suspension or aqua bidest.

Test organisms (species):

Pseudomonas putida

Details on inoculum:

TEST ORGANISMS
- Strain: Pseudomonas putida MIGULA
- extinction at 436 nm: 0.155 and 0.152

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

16 h

Remarks on exposure duration:

+/- 1 hour

Test temperature:

20.3-20.7 °C

pH:

Stock solution concentration pH before adjustment pH after adjustment
12.5 g/l 11.97 7.38
8.75 g/l 11.84 7.50

Nominal and measured concentrations:

1750; 2500; 3500; 5000; 7000; 10000 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Renewal of test solution: no
- Exposure vessel type: Sterile 250 ml Erlenmeyer flasks with cellulose  plugs
- Number of replicates: 2 independent tests with each 3 replicates per exposure concentration; 5 growth controls (no test substance); 6 sterile 
controls (no test organisms)

TEST PARAMETER
- light absorption at 436 nm, determined with Zeiss photometer PM 6: test solutions vs. sterile controls;  growth controls vs. deionized water

STATISTICAL METHODS: Multiplan computer program "MIGULA" (regression analysis)

Reference substance (positive control):

no

Duration:

16 h

Dose descriptor:

EC10

Effect conc.:

1 469 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: mean value

Duration:

16 h

Dose descriptor:

EC50

Effect conc.:

> 10 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: mean value

Details on results:

Test 1: EC10 = 1697 mg/l,  EC50 > 10 g/l
Test 2: EC10 = 1241 mg/l,  EC50 > 10 g/l

Validity criteria fulfilled:

not specified

Conclusions:

The EC50 (16 hours) was determined as >10 g/L indicating that the test substance is not harmful to aquatic microorganisms.

Executive summary:

The effects of the test item on the bacteria species Pseudomonas putida were investigated by Hüls AG (1993) according to DIN 38412, part 8 (national German guideline). Six concentrations + control were tested over a period of 16 hours. The pH of the stock solutions was adjusted to achieve a neutral value.

The EC50 was determined as >10 g/L and the EC10 as 1469 mg/L.

The study was assessed as "reliable without restrictions".

Description of key information

The effects of the substance on the bacteria species Pseudomonas putida were investigated by Hüls AG (1993) according to DIN 38412, part 8 (national German guideline). Six concentrations + control were tested over a period of 16 hours. Two tests with and without pH adjustment were performed. 
The EC50 for the test without pH adjustment was determined as 89 mg/L and the EC10 as 72 mg/L indicating that the test substance may be harmful to aquatic microorganisms.
Due to the basic properties of the test substance the concentrations used in the first test were rather low. Another test  with higher concentrations and was conducted and the pH of the stock solutions was adjusted to achieve a neutral value. The EC50 was determined as >10 g/L and the EC10 as 1469 mg/L.

Key value for chemical safety assessment

EC50 for microorganisms:

89 mg/L

EC10 or NOEC for microorganisms:

72 mg/L