Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Start date 14/05/84, finish date 18/05/84, report date 16/07/84
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP compliant and OECD equivalent. Based on nominal and measured concentrations but no supporting analytical data.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1984
Report Date:
1984

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
not specified
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): DEFI
- Analytical purity: 66% (sodium acyl isethionate)
- Impurities (identity and concentrations): Free fatty acid (21%), sodium isethionate (7%)
- Composition of test material, percentage of components:
sodium acyl isethionate (66%)
Free fatty acid (21%)
sodium isethionate (7%)
water (2%)
misc. (4%)
- Purity test date: 1983-01-31
- Stability under test conditions: Stabillty of solid is good, can be stored at room temperature. In solution, at room temperature and neutral pH, solutions are stable, at hlgh temperature (greater than 50 degrees C) and low pH hydrolysis will occur to some extent.
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
Not applicable

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Concentrations of certain selected test solutions were determined at 0, 48 and 96 hours according to the methylene blue method , however because of precipitation problems, additional pre-analytical treatment was required for some samples in order to obtain a satisfactory determination, i.e. the 48 hr (old) samples were first analysed with precipitate present and then analysed again after the additional solubilising treatment.
The temperature, pH, dissolved oxygen and total hardness (as mg/l CaC03) of the analysed test solution concentrations were also measured.

See appendex 1!!

Test solutions

Vehicle:
yes
Details on test solutions:
The test solutions are prepared by adding the appropriate amount of stock solution to each tank by pipette or measuring cylinder depending on volume and filling up to about 1" below the 10 litre mark with URPSL carbon filtered tap water, (renewal solutions are made up to the 10 litre mark at
once), the missing volume allows for addition of the fish in 500 mls -1 litre before final volumes are made up without dilution of the test
liquors.

Both sets of test vessels are labelled and when the 0 hour test solutions are prepared they are left covered and aerating on the shelves in the temperature controlled room. The temperature of the starvation tank and the test solutions are checked.

Test organisms

Test organisms (species):
Oncorhynchus mykiss (previous name: Salmo gairdneri)
Details on test organisms:
TEST ORGANISM
- Common name: Rainbow trout
- Source: Chirk fish Hatchery, Nr Wrexham 29/03/84
- Age at study initiation (mean and range, SD): ~4 months
- Feeding during test
- Food type: commercial trout food
- Amount: equivalent to about 5% total bodyweight
- Frequency: 4-5 times a day


ACCLIMATION
- Acclimation period: 29/03/84- 09/04/84
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: see above
- Feeding frequency: see above
- Health during acclimation (any mortality observed): The cumulative mortality during the acclimation period was high (14%) but fell to 2.8% over the 7 day period 21.4.84 to 28.4.84. The mortality rate is therefore well below the maximum permissab1e value of 5% over 7 days.


Study design

Test type:
semi-static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
Not applicable

Test conditions

Hardness:
118+/-8 mg/l CaCO3
Test temperature:
12.2+/-0.6 DEGREES C
pH:
8.0+/-0.2
Dissolved oxygen:
9.2+/- 0.8 mg/L
Salinity:
N/A
Nominal and measured concentrations:
Nominal 0.0*, 3.2*, 5.6, 10.0*, 18.0, 32.0, 100.0* mg/L
Measured: see table below
Details on test conditions:
TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): CLOSED
- Material, size, headspace, fill volume: 10L, GLASS
- Aeration: Continuous
- Type of flow-through (e.g. peristaltic or proportional diluter): none
- Renewal rate of test solution (frequency/flow rate): every 48hrs
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2 vessels per concentration
- No. of vessels per control (replicates): 2 vessels
- No. of vessels per vehicle control (replicates): same as control
- Biomass loading rate: no data


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Unilever carbon filtered tap water
- Total organic carbon: N/A
- Particulate matter: N/A
- Metals: NO DATA
- Pesticides: NO DATA
- Chlorine: NO DATA
- Alkalinity: NO DATA
- Ca/mg ratio: NO DATA
- Conductivity: NO DATA
- Culture medium different from test medium: NO
- Intervals of water quality measurement: 0 , 48, 96 Hrs


OTHER TEST CONDITIONS
- Adjustment of pH: NO
- Photoperiod: 16 hrs
- Light intensity: NO DATA


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality


TEST CONCENTRATIONS
- Spacing factor for test concentrations: no data
- Justification for using less concentrations than requested by guideline: N/A
- Range finding study N/A

Reference substance (positive control):
no

Results and discussion

Effect concentrations
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
> 25 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat.
Basis for effect:
mortality
Remarks on result:
other: NO DATA
Details on results:
There were no deaths in any of the concentrations except in the highest (100 mg/L. ) where only 10% (2 out 10 in tank 14) mortality occurred. The 96 hour LC50 of Defi to 4 month old feeding rainbow trout fry must therefore be greater than the maximum concentration remaining in the test liquor after 96 hours. This concentration is approximately 25 mg/L.
Results with reference substance (positive control):
Not applicable
Reported statistics and error estimates:
Statistical analysis of percentage mortality data will be performed by the methods put forward by Stephan 1977. Three methods of calculating the LC50 will be used:-
(1) Binomial ± 95% confidence limits
(2) Moving average ± 95% confidence limits
(3) Probit ± 95% confidence limits.

Applicant's summary and conclusion

Conclusions:
Analytical results indicate that Defi is rapidly lost from the test liquor. As foaming was not observed in any tank and as the additional pre-treatment
made the remaining test compound available for analysis, it is most probable that the compound was biodegraded.

There were no deaths in any of the concentrations except in the highest (100 mg/L. ) where only 10% mortality occurred. The 96 hour LC50 of Defi
to 4 month old feeding rainbow trout fry must therefore be greater than the maximum concentration remaining in the test liquor after 96 hours.
This concentration is approximately 25-30 mg/L
Executive summary:

Analytical results indicate that Defi is rapidly lost from the test liquor. As foaming was not observed in any tank and as the additional pre-treatment made the remaining test compound available for analysis, it is most probable that the compound was biodegraded.

There were no deaths in any of the concentrations except in the highest (100 mg/L. ) where only 10% mortality occurred. The 96 hour LC50 of Defi to 4 month old feeding rainbow trout fry must therefore be greater than the maximum concentration remaining in the test liquor after 96 hours. This concentration is approximately 25-30 mg/L