Bis(3,5-bis(1,1-dimethylethyl)-2-hydroxybenzoato-O1,O2)zinc

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

26 April to 30 April 1988

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study report is conducted to OECD and EEC guidelines and contains a statement of GLP compliance

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

84/449/EEC

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Bis(3,5-bis(1,1-dimethylethyl)-2-hydroxybenzoato-O1,O2)zinc

EC Number:

403-360-0

EC Name:

Bis(3,5-bis(1,1-dimethylethyl)-2-hydroxybenzoato-O1,O2)zinc

Cas Number:

42405-40-3

Molecular formula:

C30H42O6Zn

IUPAC Name:

bis(3,5-bis(1,1-dimethylethyl)-2-hydroxybenzoato-O1,O2)zinc

Details on test material:

- Physical state: white powder
- Analytical purity: not known
- Impurities (identity and concentrations): not known
- Composition of test material, percentage of components: not known
- Lot/batch No.: not known
- Expiration date of the lot/batch: not known
- Storage condition of test material: in a glass container at room temperature in the dark
- Other:

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0.025 - 0.25 μg/mlof test substance in a methanol/ distilled water (50:50) dilution solution.
- Sampling method: 10ml sample made upto volume with methanol/ distilled water (50:50)

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
2.0 g of test material were stirred into 5 L of de-ionised water overnight at room temperature. The saturated solution was then filtered through glass fibre paper and diluted to give the anticipated test concentrations 111, 200, 356, 622, 1111 and 2000 ml made upto a final volume of 20 litres. Actual test concentrations were confirmed via chemical analysis.

STABILITY OF TEST SOLUTIONS:
Ensured by daily renewal of the test media and verified by chemical analysis at 0, 24 and 96 hours.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout
- Source: Parkwood Trout Farm, Kent UK
- Length at study initiation (length definition, mean, range and SD): 5.0cm (SD = 0.4cm)
- Weight at study initiation (mean and range, SD): 1.72 g (SD = 0.28g)
- Feeding during test: none


ACCLIMATION
- Acclimation period: 8 days
- Acclimation conditions (same as test or not): same as test
- Type and amount of food: Commercial trout pellets, discontinued 24 hours prior to test
- Health during acclimation (any mortality observed): Mortality: <1%

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported

Test conditions

Hardness:

Total Hardness ≈ 350 mg/l as calcium carbonate. Although slightly higher than the recommended range for this test, it is not considered to have had any significant effects on the results of the test.

Test temperature:

14 ±1C

pH:

pH: 8.2 - 8.5

Dissolved oxygen:

>9.6 mgO2/l

Salinity:

Sodium content: 43.8 mg/l

Nominal and measured concentrations:

Anticipated test concentrations: 1.0, 1.8, 3.2, 5.6, 10 and 18 mg/l
Measured test concentrations: 0.76, 1.3, 2.5, 4.6, 7.7 and 14 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria holding 20 l of test media
- Renewal rate of test solution (frequency/flow rate): Medium renewed daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates):two
- No. of vessels per control (replicates): two
- Biomass loading rate: 0.86 g bodyweight/ litre


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised laboratory water
- Total organic carbon: mean 4.52 mg/l
- Particulate matter: No information
- Metals: No information
- Pesticides: No information
- Chlorine: mean 0.09 mg/l
- Alkalinity: mean 215.22 mg/l (calcium carbonate)
- Ca/mg ratio: mean Ca: Mg ratio = 16:1
- Conductivity: mean 905.31
- Culture medium different from test medium: No information
- Intervals of water quality measurement: 3 monthly intervals.


OTHER TEST CONDITIONS
- Adjustment of pH: no adjustment
- Photoperiod: 16 h light : 8 h dark
- Light intensity: no information


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Observations made at intervals of 3, 6, 24, 48, 72 and 96 hours

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.5 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 4.3 - 7.0 mg/l

Details on results:

- Behavioural abnormalities: Increased pigmentation at dose levels 4.6, 7.7 and 14 mg/l plus lethargy at the two highest dose levels
- Observations on body length and weight: no information
- Other biological observations: no further observations reported
- Mortality of control: None
- Other adverse effects control: None reported
- Abnormal responses: None reported
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None reported
- Effect concentrations exceeding solubility of substance in test medium: None reported

Results with reference substance (positive control):

No positive control data are included

Reported statistics and error estimates:

None reported

Any other information on results incl. tables

Sublethal observations / clinical signs:

Time (h)	LC 50 (mg/l)	95% Confidence Limits (mg/l)
3	>14	-
6	>14	-
24	11	9.3 - 14
48	7.4	6.0 - 9.2
72	7.0	5.8 - 8.5
96	5.5	4.3 - 7.0

The LC50 values were calculated from the 0 hour measured test concentrations since the material has been shown to be stable in water over a 24 hour period. Chemical analysis was not conducted on the '18 mg/l' concentration since, routinely, only five test concentrations were analysed. However, the same test solution was used in the concurrent acute toxicity to Daphnia study and, for this study, a value of 14 mg/l was determined by chemical analysis. This value is identical to that obtained by calculation based on known concentrations of the resulting test series for the acute toxicity to trout test.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96 hour LD50 to fish of the test substance is 5.5 mg/l

Executive summary:

The acute toxcity to fish study was conducted with rainbow trout under semi-static conditions and with analysis provided by HPLC with a UV-visible detector. Conducted over a 96-hour exposure period, the test substance achieved a LD₅₀ of 5.5 mg/l. On the basis of this result, the substance is considered to be classified as toxic to aquatic organisms.