oils, palm, propoxylated

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Endpoint:

toxicity to microorganisms, other

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21/09/1992 to 24/09/1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

The study of bacterial toxicity, based on oxygen consumption assessment, was performed accroding to a method developed by Huls AG (Dr Schoberl, the author).
The principle of the method was based on the mixing of a bacterial culture solution with the test substance, at a range of concentrations for 5.5 hours at 25˚C in a shaking incubator. The difference between the oxygen content in the vessels at time zero (0) and after the Incubation time was used to determine the bacterial oxygen consumption.

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Common name: Marlosoft RP
- Source and lot/batch No.of test material: 40050951
- Date of manufacture: 21/05/1992
- Expiration date of the lot/batch: <1 year (at room temperature)
- Purity test date: Not stated
- Purity: C: 73.14%; H: 11.20%; O: 14.74%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: not stated
- Stability under test conditions: <1 year at room temperature
- Solubility and stability of the test substance in the solvent/vehicle: not stated
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: not stated

Analytical monitoring:

no

Details on sampling:

Limited information provided

Vehicle:

yes

Remarks:

emulsifier

Details on test solutions:

Emulsifier: nonylphenolethoxyproproxylat (10 moles of ethylene and 5 mol of propylene oxide) was added to the test flask after the test substance was added and before the test medium was added.

Test organisms (species):

Pseudomonas putida

Details on inoculum:

- Source of culture: Institute for Water, Soil and Air Hygiene of the BGA (Berlin).
- Culture details: Liquid culture, approximately 19 hours incubation, count approx 10 x 10 5 /ml.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

5.5 h

Hardness:

Not stated

Test temperature:

23.9 to 26.9˚C

pH:

Not stated

Dissolved oxygen:

Not stated

Salinity:

Not stated

Conductivity:

Not stated

Nominal and measured concentrations:

The laboratory added 10, 50, 100 or 200 µL per bottle, resulting in concentrations of 0.1, 0.5, 1 and 2 mL/L (or 0.01, 0.05, 0.1, 0.2%), respectively. Control was prepared without the addition of the palm oil. The molecular weight of the test substance is not provided and therefor it is not possible to calculate

Details on test conditions:

TEST SYSTEM
- Test vessel: 100ml BOD bottles with stoppers.
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: not stated
- Aeration: stirred with magnetic stirrer.
- No. of vessels per concentration (replicates): 4 flasks per concentration
- No. of vessels per control (replicates): 4 flasks without test concentration, but with added HgClz, for determination of the starting O2 content.
- No. of vessels per control (replicates): 5 flasks without test substance and without HgClZ additive.
- No. of vessels per abiotic control (replicates): abiotic control included, in which HgClZ solution was added to the flasks to kill the bacteria.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: A 5 litre steep breasted bottle with a magnetic stirrer and 3520ml of double distilled water, a siphon and a venrtiliation device was coverered with aluminium foil and autoclaved at 120˚C, at 1 bar. After cooling to room temperature, 200ml of sterlie solution I and sterile stock solution II (see below) were added and the solution stirred for 30 minutes whilst stirred.
- Inoculation concentrations: 80ml of bacterial suspension added to all test vessels, count approx 10 x 10 5 /ml.

OTHER TEST CONDITIONS
- Adjustment of pH: not stated
- Photoperiod: not stated
- Light intensity: not stated
- Details on termination of incubation: not stated
- Incubtation: shaken in an incubator
- Stopage of the reaction: at the end of the test 1 ml of HgCl2 solution was added to the flasks to stop the biochemical reaction before measurement of the oxygen concentrations.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : oxygen consumption

TEST CONCENTRATIONS
- Range finding study : a preliminary study was carried out to determine the concentrations for the final test.

Reference substance (positive control):

not specified

Key result

Duration:

5.5 h

Dose descriptor:

EC10

Effect conc.:

> 2 other: ml/L

Nominal / measured:

nominal

Remarks on result:

other: Trial SK-92/10.1

Key result

Duration:

5.5 h

Dose descriptor:

EC10

Effect conc.:

> 2 other: ml/L

Nominal / measured:

nominal

Remarks on result:

other: SK-92/10.2

Details on results:

No toxicity observed in the toxicity controls.

Results with reference substance (positive control):

Not applicable

Table 1. Results of run 1

	Test substance initial concentrations
	Control	0.1 ml/L	0.5ml/L	1ml/L	2ml/L
Samples with HgCl2 – O2 initial values	7.98 7.83 7.90 8.04	8.06 8.07	7.95 7.97	7.79 8.01	7.43 7.71
Average O2 initial value	7.94	8.07	7.96	7.90	7.57
Difference between 7.94 and initial value		-0.13	-0.02	0.04	0.37
Samples without HgCl2 – O2 – end values	3.35 3.02 2.81 3.05	1.65 1.81	1.40 1.60	1.35 1.63	1.11 1.25
Average O2 end value	3.06	1.73	1.5	1.49	1.18
O2 consumption during incubation	4.88	6.34	6.46	6.41	6.39
Difference between control and samples (O2 consumption during incubation)		-1.46	-1.58	-1.53	-1.51
% effect of incubation		-29.8	>32.4	-31.4	-30.9

Table 2. Results of run 2.

	Test substance initial concentrations
	Control	0.1 ml/L	0.5ml/L	1ml/L	2ml/L
Samples with HgCl2 – O2 initial values	8.04 8.12 7.99 7.99	8.25 8.01	8.05 8.09	7.99 8.00	7.85 7.75
Average O2 initial value	8.04	8.13	8.07	8.00	7.80
Difference between 8.04 and initial value		-0.09	-0.04	0.04	0.24
Samples without HgCl2 – O2 – end values	4.16 4.52 4.12 4.20	2.95 3.10	2.70 2.66	2.66 2.91	2.44 2.52
Average O2 end value	4.25	3.03	2.82	2.79	2.48
O2 consumption during incubation	3.79	5.11	5.26	5.21	5.32
Difference between control and samples (O2 consumption during incubation)		-1.32	-1.47	-1.43	-1.54
% effect of incubation		-34.9	-38.8	-37.6	-40.6

Validity criteria fulfilled:

not specified

Conclusions:

The EC10 of propoxylated palm oil to the test organism (Pseudomonas putida) was determined at >2ml/L, although the test substance is not toxic to aquatic organisms at the limit of solubility.. No conversion of this to mg/L was possible.

Executive summary:

The test organism (Pseudomonas putida) was used to determine bacterial toxicity via an oxygen consumption inhibition test. The bacterial culture was incubated with the propoxylated palm oil at various concentrations in 100-mL “Biochemical Oxygen Demand” (BSB in German) bottle for 5.5 hours at 25 ± 2 °C. This method was not according to a guideline, however it was conducted to GLP principles.

 

As propoxylated palm oil is a liquid, concentrations were given in mL/L. Concentrations of 0.1, 0.5, 1 and 2 mL/L (or 0.01, 0.05, 0.1, 0.2% respectively) were used. A control was prepared without the addition of the palm oil.

 

All four tested concentrations resulted in an increase in oxygen consumption compared to control, indicating there was no bacterial toxicity (i.e. no inhibition of oxygen consumption) upon exposure to propoxylated palm oil.

 

The laboratory provided the EC10 for both experiments of > 2 mL/L, although the test substance is not toxic to aquatic organisms at the limit of solubility. No conversion of the units to mg/L was possible based on the information provided.

Description of key information

The EC10 of propoxylated palm oil to the test organism (Pseudomonas putida) was determined at >2ml/L, although the test substance is not toxic to aquatic organisms at the limit of solubility. No conversion of this to mg/L was possible.  

Additional information

The test organism (Pseudomonas putida) was used to determine bacterial toxicity via an oxygen consumption inhibition test. The bacterial culture was incubated with the propoxylated palm oil at various concentrations in 100-mL “Biochemical Oxygen Demand” (BSB in German) bottle for 5.5 hours at 25 ± 2 °C. This method was not according to a guideline, however it was conducted to GLP principals.

 

As propoxylated palm oil is a liquid, concentrations were given in mL/L. Concentrations of 0.1, 0.5, 1 and 2 mL/L (or 0.01, 0.05, 0.1, 0.2% respectively) were used. A control was prepared without the addition of the palm oil.

 

All four tested concentrations resulted in an increase in oxygen consumption compared to control, indicating there was no bacterial toxicity (i.e. no inhibition of oxygen consumption) upon exposure to propoxylated palm oil.