Registration Dossier

Administrative data

Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1980-05-20 to 1980-11-09
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Good quality study to GLP; considered adequate for assessment

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1983
Report Date:
1983
Reference Type:
secondary source
Title:
Unnamed
Year:
2000

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Toxicity of a C10-12 chlorinated paraffin (58% chlorination) to rainbow trout in a dynamic test system over a 60-day exposure period
GLP compliance:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): (i) C10-12 chlorinated paraffins, 58% chlorinated mixed with (ii) chlorinated n-undecane-6-14C; 59.1% chlorinated
- Substance type: technical product
- Physical state: unlabelled material (i) - slightly viscous clear liquid; labelled material (ii) - cloudy viscous liquid
- Analytical purity: (i) "100% chlorinated paraffins"; (ii) no data
- Impurities (identity and concentrations): (i) free hydrochloric acid (HCl) - 7 ppm; no stabiliser : (ii) - no data
- Composition of test material, percentage of components: (i) C10-12 chlorinated paraffins, 58% chlorinated mixed with (ii) chlorinated n-undecane-6-14C, 659.1% chlorinated
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: (i) R-201-198
- Expiration date of the lot/batch: (i) "dependent on routine analysis"
- Radiochemical purity (if radiolabelling): no data
- Specific activity (if radiolabelling): (ii) 1.04 mCi/g
- Locations of the label (if radiolabelling): n-undecane-6-14C
- Expiration date of radiochemical substance (if radiolabelling): no data
- Stability under test conditions: "unstable"
- Storage condition of test material: -20oC
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
none used
Radiolabelling:
yes

Sampling and analysis

Details on sampling:
- Sampling intervals/frequency for test organisms: at the end of the study
- Sampling intervals/frequency for test medium samples: radioactivity measured daily on days 1-7, then at intervals (23 times) to day 58; parent compound measured once/week for weeks 1, 2 and 3 and then once every 3 weeks
- Sample storage conditions before analysis: test medium - not stored prior to analysis;
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods):
- test medium: radioactivity measured by scintillation counting (lsc) and parent compound by solvent extraction/thin layer chromatography
- test organisms: radioactivity measured by sample oxidation/lsc and parent compound by soxhlet extraction/thin layer chromatography

Test solutions

Vehicle:
yes
Details on preparation of test solutions, spiked fish food or sediment:
PREPARATION OF SPIKED WATER
- Details of spiking: stock samples of labelled and unlabelled chlorinated paraffins prepared in acetone
- Controls: solvent only
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in final test solution: approx 500 ppm (v/v) initially

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: rainbow trout
- Strain: no data
- Source: Zeals Fish Farm, Zeals, Wiltshire, England
- Age at study initiation (mean and range, SD): "immature" + 14 days
- Length at study initiation (lenght definition, mean, range and SD): range - 33 to 60 mm
- Weight at study initiation (mean and range, SD): range - 0.54 to 3.65 g
- Weight at termination (mean and range, SD): range - 1.2 to 18.6 g
- Health status: no data
- Description of housing/holding area: All-glass aquaria (90x28x30 cm) with sliding lid- total vol. 68L
- Feeding during test: yes
- Food type: Mainstream trout food (BP Nutrition)
- Amount: "%/day of average fish weight x number of fish
- Frequency: daily

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): yes
- Health during acclimation (any mortality observed): no

Study design

Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
60 d
Total depuration duration:
0 d

Test conditions

Hardness:
34 to 71 mg/L as CaCO3
Test temperature:
Freshwater control: mean (range) 12.15 (11.8-12.8) oC
solvent control: mean (range) 11.85 (11.6-12.4) oC
pH:
Freshwater control: mean (range) 7.75 (7.40- 8.55)
solvent control: mean (range) 7.76 (7.40-8.45)
Dissolved oxygen:
Freshwater control: mean (range) 10.59 (9.9-11.3) mg/L
solvent control: mean (range) 10.71(9.9-11.3) mg/L
TOC:
2.3 to 5.9 mg/L
Salinity:
residual Cl : 15 to 35 µg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: All-glass aquarium
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 68 L fill volume
- Aeration: no
- Type of flow-through (e.g. peristaltic or proportional diluter): peristaltic
- Renewal rate of test solution (frequency/flow rate): dilution water - 500 ml/min; stock solution - 0.25 ml/min
- No. of organisms per vessel: 30
- No. of vessels per concentration (replicates): 1
- No. of vessels per control / vehicle control (replicates): 1
- Biomass loading rate: no data

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: freshwater reservoir
- Particulate matter: <1 to1.5 mg/l
- Metals (µg/L, 2 determinations): Cd (04, <0.2); Co (<0.4,<0.8); Cu (0.3,1.0); Pb (<0.6,<0.7); Mn (2,8); Ni (<0.4, 0.3); Zn (2,13); Fe (3,41)
- Pesticides: no data
- Chlorine: <4 µg/L
- Alkalinity: no data
- Ca/mg ratio: no data
- Conductance: 75-150 µs/cm
- Holding medium different from test medium: no
- Intervals of water quality measurement: weekly
- Intervals of test medium replacement: continuous

OTHER TEST CONDITIONS
- Adjustment of pH: no data
- Photoperiod: no data
- Light intensity: no data

RANGE-FINDING / PRELIMINARY STUDY : no
Nominal and measured concentrations:
Nominal : 0.032, 0.1, 0.32, 1 and 3.2 mg/L
Measured : 0.033, 0.10, 0.35, 1.07 and 3.05 mg/L
Reference substance (positive control):
no
Details on estimation of bioconcentration:
Calculated as 14C in tissues/mean exposure concentration and as parent compound in tissue/mean exposure concentration

Results and discussion

Bioaccumulation factoropen allclose all
Type:
BCF
Value:
>= 1 173 - <= 7 816
Basis:
whole body w.w.
Calculation basis:
other: concentration at day 60
Remarks on result:
other: 14C in fish
Remarks:
Conc.in environment / dose:0.033 to 3.05 mg/L
Type:
BCF
Value:
>= 574 - <= 7 273
Basis:
whole body w.w.
Calculation basis:
other: concentration at day 60
Remarks on result:
other: parent compound in fish
Remarks:
Conc.in environment / dose:0.33 to 3.05 mg/l
Details on kinetic parameters:
not calculated
Metabolites:
not examined
Results with reference substance (positive control):
not done
Details on results:
- Mortality of test organisms (out of 30): 9, 4, 17, 24 and 27 at 0.032, 0.1, 0.32, 1.0 and 3.2 mg/L nominal respectively
- Behavioural abnormalities: lethagy, failure to eat, darkening pigmentation and death. Symptoms appeared by day 3 and reached peak by day 16. Severity greater at higher concentrations
- Observations on body length and weight: Reduced weight gain seen at 1.0 and 3.2 mg/l (nominal) dose groups
- Other biological observations: none
- Organ specific bioaccumulation: no assessed
- Bound residues forming a plateau: no data
- Mortality and/or behavioural abnormalities of control: mortality - freshwater controls - 0/30, 1/30, 1/30 (series 1, 2, 3 respectively);
- Loss of test substance during test period: not applicable
- Results with vehicle control: mortality - solvent control - 0/29, 2/29, 3/29 (series 1, 2, 3 respectively)
Reported statistics:
none

Applicant's summary and conclusion

Validity criteria fulfilled:
not applicable
Conclusions:
The whole body bioaccumulation factor (BCF) for a C10-12 chlorinated paraffin (58% chlorinated, mixed with a 14C-labelled chlorinated C12 paraffin) in rainbow trout after exposure for 60 days, varied from 1173 to 7816, based on radioactivity measurement and from 574 to 7273, based on parent compound measurement.
Executive summary:

In a good-quality study conducted to GLP, the bioaccumulation factor (BCF) in rainbow trout of a C10-12 chlorinated paraffin (58% chlorination) has been determined following exposure to concentrations varying from 0.033 to 3.05 mg/L in a dynamic, flow-through system for 60 days. Groups of 30 young fish were exposed to the 58% chlorinated paraffin, mixed with 6-14C-labelled n-undecane (59.1% chlorinated), and the concentration of both radioactivity and parent compound measured in the medium during the study and in the fish at the end of the study period. The behaviour and mortality of the exposed fish was monitored during the study along with their weight and body length.

In the first series of experiments, no mortality or abnormal behaviour was noted in the control groups (freshwater and solvent control), whereas a dose-related increase in mortality (4/30, 17/29 and 24/30 at a nominal medium concentration of 0.1, 0.32 and 1.0 mg/L respecetively) was seen in the treated groups). In additional studies (series 2 and 3), mortalities were seen in all control groups (1/30 controls in series 1 and 2 and 2/29 and 3/29 solvent controls in series 2 and 3 respectively). Abnormal behaviour, inluding lethagy, failure to feed and bouts of directionless activity were seen in all treated groups, the extent being greater at the highest exposure concentrations. A 60-day LC50 value of 0.34 mg/L was determined for this material.

The BCF values, in the surviving fish, for the C10-12 chlorinated paraffin determined in this study based on radioactivity measurements were 7155, 7816, 3723, 2642 and 1173 at measured concentrations of 0.033, 0.10, 0.35, 1.07 and 3.05 mg/L respectively. The corresponding values based on parent compound measurements were 7273, 3600, 1629, 991 and 574. The general reduction in BCF with increasing exposure concentration may be related to a lack of solubility of the test material at highest concentrations.