di-, tri- and tetrachlorotetradecane

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

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:

1997

Report date:

1997

Reference 2

Reference Type:

publication

Title:

Unnamed

Year:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Deviations:

yes

Remarks:

Extended to 96 hours

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: clear liquid
- Analytical purity: 99.29%
- Impurities: 0.3% epoxy soyabean oil stabiliser
- Composition of test material, percentage of components: C14-17 chlorinated paraffin
- Lot/batch No.: 19466/01
- Purity test date: 1996-10-17
- Storage condition of test material: "in the dark in a refrigerator in a 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 of the test solutions, using the excess remaining after filling each test vessel and analysed by liquid scintillation counting. At the end of the test, each blank solution was examined in a similar manner.

Test solutions

Vehicle:

yes

Details on test solutions:

Stock solutions of radiolabelled test substance were prepared in acetone and added to sterile culture medium (0.1 ml of appropriate stock solution to 1 L medium) with vigorous stirring. All preparations appeared as homogenous dispersions suitable for testing.

Test organisms

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM- Strain: ATCC 22662- Source (laboratory, culture collection): laboratory cultures- Age of inoculum (at test initiation): 3 days- Method of cultivation: in the test mediumACCLIMATION- Acclimation period: none- Culturing media and conditions (same as test or not): same as test- Any deformed or abnormal cells observed: no data

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

extended from guideline duration of 72 h

Post exposure observation period:

none

Test conditions

Hardness:

no data

Test temperature:

daily temperatures varied from 23.9 to 24.0oC

pH:

7.4 to 7.5 at start of study and 10.0 to 10.3 at end of study

Dissolved oxygen:

no data

Salinity:

no data

Nominal and measured concentrations:

Nominal concentrations were : control (<0.025), solvent control (<0.025), 0.10, 0.18, 0.32, 0.56, 1.0, 1.8 and 3.2 mg/lMeasured concentrations at 0 h were : control (<0.025), solvent control (<0.025), 0.12, 0.17, 0.29, 0.56, 0.90, 1.5, 2.4 mg/lMeasured concentrations at 96 h were: control (<0.025), solvent control (<0.025), <0.025, <0.025, <0.025, <0.025, 0.082, 0.031, 0.056 mg/l

Details on test conditions:

TEST SYSTEM- Test vessel: 250 ml capacity borosilicate glass conical flask - Type (delete if not applicable):closed - Material, size, headspace, fill volume: 100 ml - Aeration: continuous orbital shaking- Type of flow-through (e.g. peristaltic or proportional diluter): no- Renewal rate of test solution (frequency/flow rate): no- Initial cells density: 10,000/ml- Control end cells density: control and solvent control - 520 and 589 x 10(exp)4/ml- No. of vessels per concentration (replicates): 3- No. of vessels per control (replicates): 3 - No. of vessels per vehicle control (replicates): 6GROWTH MEDIUM- Standard medium used: yesTEST MEDIUM / WATER PARAMETERSno dataOTHER TEST CONDITIONS- Sterile test conditions: yes- Adjustment of pH: no- Photoperiod: continuous illumination- Light intensity and quality: 7980 lux (measured once during study)EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : growth rate at 24 h intervals- Determination of cell concentrations: Coulter counter ZB- Chlorophyll measurement: noTEST CONCENTRATIONS- Spacing factor for test concentrations: 1.8- Range finding study : no data

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 3

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

> 3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Effect concentrations 4

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 5

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 6

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

> 3.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Details on results:

Mean area under growth curve at 96 hr (as % solvent control) for control, 0.1, 0.18, 0.32, 0.56, 1.0, 1.8 and 3.2 mg/l groups respectively: 95, 92, 84, 89, 82, 85, 84, 86% (all values statistically significantly different from solvent control except control and 0.10 mg/l)Mean growth rate at 72 hr (as % solvent control) for control, 0.1, 0.18, 0.32, 0.56, 1.0, 1.8 and 3.2 mg/l groups respectively : 101, 100, 98, 98, 97, 98, 98, 99% (values for 0.18, 0.32, 0.56 and 1.0 mg/l statistically significantly different from solvent control)

Results with reference substance (positive control):

not used

Reported statistics and error estimates:

no data

Applicant's summary and conclusion

Validity criteria fulfilled:

not applicable

Remarks:

Guidelines have no criteria for Selenastrum capricornutum

Conclusions:

Cereclor S52 (52% chlorinated) had little effect on growth of the freshwater alga, Selenastrum capricornutum, at up to a nominal concentration of 3.2 mg/l over a period of 96 h. The maximum inhibitions in growth rate was 3% and in biomass accumulation was 18%, although no clear dose-response was observed. The 96-h NOEC from this test is around 0.1 mg/l (nominal), although it should be noted that no statistically significant effects were seen at some higher concentrations, and the 96-h EC50 is >3.2 mg/l (nominal).

Executive summary:

In a OECD (201) guideline study, the effects of Cereclor S52 (C14 -17 chlorinated paraffin; 52% chlorination, containing 0.3% stabiliser) on growth of the freshwater alga Selenastrum capricornutum over a 96 h period was examined at 0 (solvent control), 0.1, 0.18, 0.32, 0.56, 1.0, 1.8 and 3.2 mg/l (nominal).

The maximum inhibition of growth, as assessed by biomass accumulation, compared with solvent controls was 18% at a nominal concentration of 0.56 mg/l, and although significant inhibition in growth was seen at nominal concentrations of 0.18 mg/l and above, there was no clear dose-response relationship.

No effects were seen on algal growth over 72 h at the two highest test concentrations (1.8 and 3.2 mg/l nominal). Small, but statistically significant, reductions in growth rate (up to 3% reduction) were seen at lower concentrations.

Although the 96-h no-observed-effect concentration (NOEC) in this study was 0.1 mg/l, it should be noted that no statistically significant effects were seen at some higher concentrations. In addition, the low level of inhibition seen means that the 96-h EC50 value is >3.2 mg/l (nominal). It is clear that little or no toxic effects were seen on algal growth in this study.