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Short-term toxicity to fish

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Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
21 January 2008-25 January 2008
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Analytical monitoring:
not specified
Details on sampling:
The water solubility of the test substance at 19.9°C ± 0.6°C was < 7 x 10·5 g/1, using the column elution method. R507-2 concentrations were not stable during freezing and thawing of samples. Consequently, samples were analysed on the day of sampling (NOTOX Project 486300).

During the final test singular samples for possible analysis were taken from the control and the undiluted filtrate according to the schedule below. The filter was also retained for possible analysis of the residue.

Frequency: at t=0 h, t=24 h and t=96 h 2ml
Volume: 2ml
Storage: Not applicable, samples were analysed on the day of sampling.

Vehicle:
not specified
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g. film of the test substance on the water surface).

The batch of R507-2 tested was a red powder with a purity ;,:96.4% and not soluble in test medium at the loading rate tested.

Preparation of test solutions started with loading rates of 100 mg/I applying a 5 to 6-minute treatment period with ultrasonic waves followed by 3 days of magnetic stirring. The resulting red dispersion contained undissolved particles and a floating layer and was consequently filtered through a 0.45 µm membrane filter (Schleicher & Schuell, RC55) to remove the larger undissolved particles. The lower test concentration was prepared by dilution of the filtrate in test medium. The final test solutions were all clear and colourless.
Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: Carp
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Length at study initiation (length definition, mean, range and SD): 2.3 ± 0.1 cm
- Weight at study initiation (mean and range, SD): : 0.41 ± 0.08 g
- Method of breeding: F1 from a single parent-pair bred in UV-treated water


ACCLIMATION
- Acclimation period: At least 12 days after delivery
- Feeding frequency during acclimation: Daily with pelleted fish food (Nutra 3.0 T, TROUW NUTRITION, Putten, The Netherlands)
- Health during acclimation (any mortality observed): In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.


FEEDING DURING TEST
No feeding from 24 hours prior to the test and during the total test period.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
96 h
Post exposure observation period:
Fish were only observed in the 96h period.
Test temperature:
21.4 - 22.8 °C.
pH:
pH of 7.6 - 7.9
Dissolved oxygen:
6.2
Nominal and measured concentrations:
Analysis of the samples taken from the filtrate prepared at 100 mg/I showed that the measured concentrations were below the limit of detection, i.e. < 0.02 mg/I. The filter residue was identified as R507-2.
Details on test conditions:
TEST SYSTEM
- Test vessel: 10 litres, all-glass, containing 9.5 litres of test solution.
- Aeration: Aeration was introduced after 24 hours of exposure.
- No. of organisms per vessel: Control and filtrate: 7 fish per test group
10% filtrate: 3 fish
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


OTHER TEST CONDITIONS
. Note that aeration was introduced after 24 hours of exposure, as the oxygen concentration tended to drop below the optimum level for testing with carp, i.e. below 5 mg/I. All test conditions remained within the ranges prescribed by the protocol (pH: 6.0-8.5, constant within 1 unit; temperature 20-24°C, constant within 2°C; oxygen> 60% of air saturation).

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
Analysis of the samples taken from the filtrate prepared at 100 mg/I showed that the measured concentrations were below the limit of detection, i.e. < 0.02 mg/I. The filter residue was identified as R507-2.
Reference substance (positive control):
yes
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Nominal / measured:
meas. (initial)
Conc. based on:
test mat. (dissolved fraction)
Basis for effect:
mortality (fish)
Details on results:
No Mortality of species was observed at any of the time points.
Results with reference substance (positive control):
During the test the pH, oxygen concentration and the temperature of the medium were within the optimal ranges for fish.

Under the conditions of the present test PCP induced no lethal effects in carp at 0.10 mg/I. The 96h-LCs0 for carp exposed to PCP was 0.20 mg/I (95 % confidence interval between 0.16 and 0.32 mg/I). This effect was already reached within 48 hours of exposure.

The range of the 96h-LC50 for carp is generally between 0.10 and 0.46 mg/I based on historical data of reference tests performed approximately every 3 months from April 1988 until the end of 2000, and annually since then. Hence, the sensitivity of carp originating from the present batch for PCP falls within the range of sensitivities generally observed during the past years.

The raw data and report from this study are kept in the NOTOX archives. The test described above was performed under GLP-conditions with a QA-check.
Sublethal observations / clinical signs:

Table1           Incidence of mortality and total mortality during the final test

 

Test group* R507-2

(%filtrate)

Initial number of fish

Cumulative mortality

Total Mortality(%)

3%h

24h

48h

72h

96h

Control

7

0

0

0

0

0

0

10

3

0

0

0

0

0

0

100

7

0

0

0

0

0

0

*Test groups represent percentages of a 0.45 µm filtered solution prepared at a loading rate of 100mg/I.

 

Table 2 shows the effect parameters based on loading rates.

 

Table2           Effect parameters

 

Parameter

Test group* R507-2

(mg/I)

NOEC

100

24,48,72,96h-LC50

>100

*Test group represents a 0.45 µm filtered solution prepared at a loading rate of 100mg/I.

Validity criteria fulfilled:
yes
Conclusions:
Under the conditions of the present test R507-2 induced no visible or lethal effects in carp when exposed to a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I (NOEC).

The 96h-LC50 exceeded the maximum solubility in test medium, i.e. the concentration present in a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I. Analyses showed that the measured concentration was below 0.02 mg/I.
Executive summary:

96-Hour Acute Toxicity Study in Carp with R507-2.

 

The study procedure described in this report was based on the OECD guideline No. 203, 1992. In addition, the procedures were designed to meet the test methods of the EEC directive 92/69, Part C.1, 1992, the ISO International Standard 7346-1: Static method, 1996 and the OECD series on testing and assessment number 23, 2000.

 

The batch of R507-2 tested was a red powder with a purity 96.4% and not soluble in test medium at the loading rate tested. The water solubility was determined to be less than 0,07 mg/I.

 

Based on the poor solubility in water it was decided to start directly with a final test. Seven carp per test group were exposed to a control and a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I. In addition, 3 carp were exposed to a ten-fold dilution of the filtrate. The total test period was 96 hours and samples for analytical confirmation of actual concentrations were taken at the start, after 24 and 96 hours of exposure.

 

Analysis of the samples taken from the filtrate prepared at 100 mg/I showed that the measured concentrations were below the limit of detection, i.e. <0.02 mg/I. The filter residue was identified as R507-2,

 

The study met the acceptability criteria prescribed by the protocol and was considered valid.

 

R507-2 induced no visible or lethal effects in carp when exposed to a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I (NOEC).

 

The 96h-LC50 exceeded the maximum solubility in test medium, i.e. the concentration present in a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I. Analyses showed that the measured concentration was below 0.02 mg/I.

Description of key information

96-Hour Acute Toxicity Study in Carp with R507-2.

 

The study procedure described in this report was based on the OECD guideline No. 203, 1992. In addition, the procedures were designed to meet the test methods of the EEC directive 92/69, Part C.1, 1992, the ISO International Standard 7346-1: Static method, 1996 and the OECD series on testing and assessment number 23, 2000.

 

The batch of R507-2 tested was a red powder with a purity 96.4% and not soluble in test medium at the loading rate tested. The water solubility was determined to be less than 0,07 mg/I.

 

Based on the poor solubility in water it was decided to start directly with a final test. Seven carp per test group were exposed to a control and a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I. In addition, 3 carp were exposed to a ten-fold dilution of the filtrate. The total test period was 96 hours and samples for analytical confirmation of actual concentrations were taken at the start, after 24 and 96 hours of exposure.

 

Analysis of the samples taken from the filtrate prepared at 100 mg/I showed that the measured concentrations were below the limit of detection, i.e. <0.02 mg/I. The filter residue was identified as R507-2,

 

The study met the acceptability criteria prescribed by the protocol and was considered valid.

 

R507-2 induced no visible or lethal effects in carp when exposed to a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I (NOEC).

 

The 96h-LC50 exceeded the maximum solubility in test medium, i.e. the concentration present in a 0.45 µm filtered solution prepared at a loading rate of 100 mg/I. Analyses showed that the measured concentration was below 0.02 mg/I.

Key value for chemical safety assessment

Additional information