di-, tri- and tetrachlorotetradecane

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996-06-25 to 1996-06-27

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study to GLP

Justification for type of information:

End points of target substance di-, tri-, tetra- chlorotetradecane can be read across from source substance Alkanes, C14-17, chloro EC number: 287-477-0 | CAS number: 85535-85-9. This due to the fact that di-, tri-, tetra- chlorotetradecane components make up the C14 content found in Alkanes, C14-17, chloro.
The degree of chlorination can greatly affect the chemistry of the substance. To reduce variability of results, studies used from the Alkanes, C14-17, chloro REACH dossier will only include studies that used C14 components and similar levels of chlorination to di-, tri-, tetra- chlorotetradecane – 35 to 52 % chlorination.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1997

Reference 2

Reference Type:

publication

Title:

Unnamed

Year:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

no data on reference substance

GLP compliance:

yes

Remarks:

UK

Test material

Test material information

Constituent 1

Reference substance name:

Alkanes, C14-17, chloro

EC Number:

287-477-0

EC Name:

Alkanes, C14-17, chloro

Cas Number:

85535-85-9

Molecular formula:

Substance is a range of chlorinated isomers of C14 to C17 paraffin

IUPAC Name:

Alkanes, C14-17, chloro

Details on test material:

- Name of test material (as cited in study report): Cereclor S52
- Substance type: technical product
- Physical state: liquid (clear)
- Analytical purity: 99.29%
- Impurities: 0.3% epoxy soyabean oil stabiliser
- Composition of test material, percentage of components: C14-17 chlorinated paraffin (52% chlorination)
- Purity test date: 1996-10-17
- Lot/batch No.: 19466/01
- Storage condition of test material: “in the dark, in a refrigerator in the container in which it was received”

Other
- Name of test material (as cited in study report): chlorinated n-pentadecane-8-14C (51% chlorinated)
- Radiochemical purity (if radiolabelling): >96.6%
- Specific activity (if radiolabelling): 39.6 MBq/g (1.07 mCi/g)
- Locations of the label (if radiolabelling): 8-14C
- Expiration date of radiochemical substance (if radiolabelling): no data
- Stability under test conditions: no data
- Storage condition of test material: frozen

Test substance: defined in the report as test material (chlorinated paraffin (52% chlorinated, C14-17)) mixed with radiolabelled material (chlorinated n pentadecane-8-14C)

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):none used

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

At the start of the test, samples were taken from each test solution using the excess remaining after filling the test vessels and at the end of the test, each solution was sampled, pooling replicates where necessary to obtain a sufficient volume. The concentration of the test substance was determined by radiochemical analysis.

Test solutions

Vehicle:

yes

Details on test solutions:

- Method: Stock solutions of the test substance were prepared in acetone. Each test solution was prepared by the addition of 0.2 ml of the appropriate stock solution to 2000 ml of dilution water. The additions were made gradually, below the surface of the medium, whilst stirring vigorously by magnetic follower.- Controls: untreated control (dilution water only), solvent control (acetone)- Chemical name of vehicle: acetone (organic solvent)- Concentration of vehicle in test medium: the solvent control and each concentration of the test substance contained 0.1 ml acetone/l- Evidence of undissolved material (e.g. precipitate, surface film, etc): no data

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM- Common name: Water flea- Strain: Straus- Source: continuous laboratory cultures- Age at study initiation (mean and range, SD): <24 hours old (first instar)- Method of breeding: diploid parthenogenesis- Feeding during test: no ACCLIMATION- Acclimation period: no data- Type and amount of food: Cultures were fed a defined diet of algae Chlorella vulgaris, strain CCAP 211/12, and a commercially available microencapsulated diet “Frippak Booster®”- Health during acclimation (any mortality observed): the test organisms and the culture from which they were obtained showed no evidence of disease before the test period

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

none

Test conditions

Hardness:

248 mg/l CaCO3

Test temperature:

20±1oC

pH:

8.01 to 8.07

Dissolved oxygen:

9.0 to 9.2 mg/l

Salinity:

freshwater

Nominal and measured concentrations:

Nominal : 0.0032, 0.0056, 0.01, 0.018, 0.032, 0.056 and 0.1 mg/lMeasured (0 and 48 hours): 0.0025, 0.0041, 0.0094, 0.015, 0.024, 0.047, 0.095 mg/l

Details on test conditions:

TEST SYSTEM- Test vessel: borosilicate glass beakers of 250 ml nominal capacity - Type (delete if not applicable): closed - Material, size, headspace, fill volume: 200 ml test solution, depth ~60 mm - Aeration: no- No. of organisms per vessel: 5- No. of vessels per concentration (replicates): 4- No. of vessels per control (replicates): 4- No. of vessels per vehicle control (replicates): 4- Biomass loading rate: no dataTEST MEDIUM / WATER PARAMETERS- Source/preparation of dilution water:reconstituted water medium (Elendt’s M4 Daphnia medium), aerated for >2 hours before use- Particulate matter:- total organic carbon: 1.6 mg/l (non-purgable)- Metals: cadmium <1 µg/l, copper 1 µg/l, cobalt <1 µg/l, iron 29 µg/l, lead 2 µg/l, manganese 65 µg/l, nickel <1 µg/l, zinc 1 µg/l- Alkalinity: 48.8 mg/l as CaCO3- Conductivity: 663µS/cm- Culture medium different from test medium: no- Intervals of water quality measurement: onceOTHER TEST CONDITIONS- Photoperiod: 16 hours light/8 hours dark, with 20 min transition periodsEFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Each Daphnia was viewed by eye, and was defined as affected if showing no whole body movement relative to the water within a period of 15 seconds, even if movement of the appendages was visible. Daphnia so affected were termed immobile. Any overt symptoms of toxicity were also recorded. Observations were made at 24 and 48 hours after commencing the test.TEST CONCENTRATIONS- Spacing factor for test concentrations: 1.8- Range finding study: no

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.008 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test material mixed with 14C-labelled chlorinated pentadecane

Basis for effect:

mobility

Remarks on result:

other: 0.0059-0.0097 mg/l

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.006 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

other: test material mixed with 14C-labelled chlorinated pentadecane

Basis for effect:

mobility

Remarks on result:

other: 0.0047-0.0073 mg/l

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

other: test material mixed with 14C-labelled chlorinated pentadecane

Basis for effect:

mobility

Effect concentrations 4

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 0.095 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

other: test material mixed with 14C-labelled chlorinated pentadecane

Basis for effect:

mobility

Details on results:

Immobilisation at 24 h- control, solvent control, 0.0032, 0.0056 and 0.01 mg/l (nominal): 0/20 immobilised- 0.018, 0.032, 0.056 and 0.1 mg/L (nominal): 5/20, 6/20, 8/20, 7/20 immobilised respectivelyImmobilisation at 48 h- control, solvent control, 0.0032 mg/l (nominal): 0/20 immobilised- 0.0056, 0.01, 0.018 and 0.032 mg/l (nominal): 9/20, 18/20, 15/20 and 17/20 immobilised- 0.056 and 0.1 mg/l (nominal): 20/20 immobilisedOther observations:- a number of animals exposed to each of the test substance treatments displayed a red coloration on parts of the exoskeleton.

Results with reference substance (positive control):

no data

Reported statistics and error estimates:

The 24 and 48 hour EC50 values, and their 95% confidence intervals, were calculated by computer program, using Stephan’s method.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

<10% immobilisation in controls ; dissolved oxygen concentration >3 mg/l

Conclusions:

Exposure of Daphnia magna to Cereclor S52 for 48 hours in freshwater results in substantial immobilisation at concentrations of 0.0056 mg/l and above, with complete immobilisation at concentrations of 0.056 mg/l and greater. The 48-hr EC50 value of 0.0077 mg/l (nominal concentration) or 0.0059 mg/l (measured concentration), indicates that the material is very toxic to aquatic invertebrates in this test system

Executive summary:

A Daphnia magna acute immobilisation test was performed with Cereclor S52 (a C14-17 chlorinated paraffin; 52% chlorinated, containing 0.3% stabiliser) in freshwater under static conditions, conducted according to OECD test guideline 202. The test substance was mixed with radiolabelled chlorinated n-pentadecane-8-¹⁴C (51% chlorinated) to facilitate analysis of test solution concentrations, and dissolved in acetone to prepare the necessary dilutions since it had low aqueous solubility. The test material was examined up to a concentration of 0.1 mg/ml (nominal). Although analytical concentrations of the test substance were only 40 to 79% of nominal at the end of the study, the test was valid by OECD criteria (not more than 10% immobilisation was seen in controls and dissolved oxygen concentration at the end of the test was greater than 3 mg/l). No reference substance (positive control) results were presented; however, there is no indication that the test conditions were unreliable. The Daphnia were not fed during the test.

 

No immobilisation was seen in untreated or solvent controls or at the lower concentrations of the test substance (up to 0.01 mg/l) after 24 hours exposure and less than 40% immobilisation was observed at the higher concentrations (0.018-0.1 mg/l). Therefore it was not possible to calculate an EC50 for this timepoint. There was no immobilisation in the untreated or solvent controls or at the lowest concentration of the test substance (0.0032 mg/l) after 48 hours exposure; however immobilisation rates were 9/20, 18/20, 15/20, 17/20 for the four intermediate concentrations respectively (0.0056, 0.01, 0.018, 0.032 mg/l), and there was complete immobilisation at the two highest concentrations (0.056-0.1 mg/l). The 48-hour EC50 was calculated to be (nominal) 0.0077 mg/l (95% confidence limits 0.0059 to 0.0097 mg/l).