Amines, C12-14-alkyl

Sediment toxicity

Administrative data

Endpoint:

sediment toxicity: long-term

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication/study report which meets basic scientific principles.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2000

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

The toxicity of the test substance to the nematode Caenorhabditis elegans was determined in artificial sediment spiked in various concentrations. The sediment bioassay was carried out according to Traunspurger et al. 1997. Test under ISO standardisation.
(Traunspurger W., Haitzer M., Höss S., Beier S., Ahlf W. & Steinberg C. (1997) Ecotoxicological assessment of aquatic sediments with Caenorhabditis elegans (nematoda) - A method for testing in liquid medium and whole sediment samples. Environmental Toxicology and Chemistry, 16, 245-250.)

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Amines, tallow alkyl

EC Number:

263-125-1

EC Name:

Amines, tallow alkyl

Cas Number:

61790-33-8

IUPAC Name:

61790-33-8

Constituent 2

Reference substance name:

Amines, tallow alkyl

IUPAC Name:

Amines, tallow alkyl

Details on test material:

- Name of test material (as cited in study report): Tallow alkylamine, Genamin TA 100 D

Sampling and analysis

Analytical monitoring:

no

Test substrate

Vehicle:

yes

Details on sediment and application:

PREPARATION OF SPIKED SEDIMENT
- Pooling or mixing of different substrates: artificial micxed sediment with 44% sand, 48% silt and 8% clay
- Details on spiking: Tallow alkylamine was dissolved in pure ethanol. 10 µL of the ethonol stock solution was mixed thoroughly with 0.5 g sediment (wet weight; 26% water content) that was already mixed with 0.5 ml of M9-medium (phosphate buffer) in the test vessels (Nunc polystyrol multiwells). Thus, the maximum ethanol concentration did not exceed 1 % which is tolerated by the nematodes. In order to allow the chemicals to equally distribute between aqueous and solid phase, the spiked sediment was incubated for 24 h before the start of the test.

Test organisms

Test organisms (species):

Caenorhabditis elegans

Details on test organisms:

TEST ORGANISM
- Cultivation and storage: C. elegans is maintained on agar plates ( 10 cm petri dishes with NGM-agar) with a bacterial lawn (E. coli OP 50). When bacteria are used up, C. elegans forms dauer larvae due to lack of food. These so called "starved plates“ serve as stock cultures for C. elegans that can be stored at least for two months. E.coli are stored as glycerin stock cultures at –20 °C.

-Preparation of test organisms: When synchronous worms are needed for a test, dauer larvae are transferred on an agar plate with a fresh lawn of E. coli. After about three days at 20°C a lot of gravid hermaphrodites, as well as juveniles of stage 1 and 2, are found on the plate. The test starts with worms in the first larval stage. In order to obtain worms synchronized to this life stage, the plates are rinsed with M9-medium. The suspension containing
the nematodes is then filtered through a nylon net of 10 Fm mesh and subsequently through a net of 5 Fm mesh size to retain larger juveniles and adults. The filtered suspension contains only first stage juveniles. The length of these individuals was measured as 270Fm + 16Fm (N=50).

- Age of animals at beginning of exposure: juvenile worms of the first stage (J1)

Study design

Study type:

laboratory study

Test type:

static

Water media type:

freshwater

Type of sediment:

artificial sediment

Limit test:

no

Exposure duration

Duration:

72 h

Exposure phase:

total exposure duration

Test conditions

Test temperature:

20°C

Nominal and measured concentrations:

Nominal concentrations: 1350 mg/kg (dw) + three lower concentration not exactly mentioned in test report, highest concentration 2030 mg/kg (dw) where the limit of solubility was reached

Details on test conditions:

Before the start of the assay, 0.25 ml of a bacterial suspension (Escherichia coli in M9-medium) were added to each test vessel as food for the nematodes. After that 10 juvenile worms of the first stage (J1) were added to each test vessel, containing now 0.5 g sediment (ww), 0.5 ml test solution and 0.25 ml bacterial suspension. The vessels were then incubated for 72 h on a shaker at 20 °C. Five (three) replicates were set up for the main test (range finding test). In order to stop the assay, the nematodes were heat killed and stained with Rose Bengal. After extracting the nematodes from the sediment (centrifugation in a density gradient, using Ludox T 50), body length and number of eggs inside the body were determined under a microscope.


CHARACTERIZATION OF ARTIFICIAL SEDIMENT
- Particle size distribution
- % sand: 44
- % silt: 48
- % clay: 8
- Composition (if artificial substrate): Quarz Millisil W4 (Quarzsandwerke GmbH), Calzitic sand (Pitti Heimtierprodukte, Germany), Kaolin (Sigma, K 7375), Dolomit 5T (Carl Jäger Tonindustriebedarf GmbH), Humus (Sphagnum peat; Terra Plant service Analyse & Consulting GmbH), Eisenoxid(III)-oxid red (Fluka 44956), Aluminum-III-oxide (Riedel de Haen)
- Moisture: 26% water content


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : growth, egg production, fertility


TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Range finding study
- Test concentrations: 135, 500, 1350 mg/kg sed (dw)
- Results used to determine the conditions for the definitive study: yes

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 2 030 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

>= 2 030 mg/kg sediment dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Any other information on results incl. tables

For this substance we did not find an effect up to the highest tested concentration of 2030 mg/kg Sed (dw). Here we reached the limit of solubility. When the stock solution was added to the sediment, a white precipitate occurred at high concentrations.

Applicant's summary and conclusion

Executive summary:

Body length, eggs per worm and % of gravid worms are shown in Figure 1. Threshold values (NOEC, LOEC) are given in Table 1. Mean values of the single replicates are shown in Appendix B (Table I-IV). Sodium tetrapropenyl benzene sulfonate: The range finding test showed that there was no effect at 135 mg/Kg Sed (dw), whereas we found a total inhibition of all test parameters at 500 mg/Kg Sed (dw). In order to render NOEC and LOEC more precisely, smaller concentration steps were investigated in the main test. The results of the main test agreed well with those of the range finding test. The NOEC was found at 100, the LOEC at 200 mg/Kg Sed (dw). Cocos dimethylamine: The results of the range finding test and the main test are slightly different, however they are in the same order of magnitude (Fig. 1). We took the results of the main test for calculations of NOEC and LOEC, because of the higher number of replicates and thus higher statistical significance. Di hardened tallow dimethyl ammonium chloride: Here we did not find an effect in the range finding test up to the highest tested concentration of 1350 mg/Kg Sed (dw). Thus we investigated higher concentrations (up to 5410 mg/Kg Sed (dw)) in the main test. The NOEC-value was found at 1350, the LOEC-value at 2030 mg/Kg Sed (dw). Tallow alkylamine: For this substance we did not find an effect up to the highest tested concentration of 2030 mg/Kg Sed (dw). Here we reached the limit of solubility. When the stock solution was added to the sediment, a white precipitate occurred at high concentrations.