Cyclohexane-1,2,4-triyltris(ethylene)

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP-conform guideline study with acceptable limitations. The anaylitcal method is not substance-specific (measurement of TOC). The initial loading of the saturated solution is not specified in the description of the test procedure.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

(1984)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

EC 92/69 EWG part C1

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: DIN 38412 Part 1

Principles of method if other than guideline:

Method: EEC Guideline 92/69/EEC C1 and OECD guideline No.203

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: A saturated solution has been tested and was diluted to a dilution level of 1/1, 1/2, 1/4, 1/8 and 1/16.
- Sampling method: Analysis of total organic carbon

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A saturated solution of the test substance in drinking water has been prepared and was stirred 18 h. After that the suspension has been filtrated and has been used for the test by preparation of a dilution series. The solution was diluted with a factor of 2 to dilution levels of 1/16 ; 1/8 ; 1/4 ; 1/2, and 1/1 of the saturated solution.
- Differential loading: dilution levels of 1/16 ; 1/8 ; 1/4 ; 1/2, and 1/1 of the saturated solution
- Controls: water without test substance
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): not specified

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebra fish
- Strain: Brachydanio rerio
- Source: West Aquarium, Bad Lauterberg
- Weight at study initiation (mean and range, SD): 0.37 g
- No feedung during test

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions (same as test or not): Maintenance in aquaria with 200 L (stone aquaria) or 300 L (glass). Pretreatment with malachite green, 3-times per week, 14 days quarantine
- Type and amount of food during acclimation: TetraMin, 1% of body weight
- Feeding frequency during acclimation: daily
- Health during acclimation (any mortality observed): at test begin the fish showed normal behaviour and no signs of diseases.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

13.0 °dH

Test temperature:

20 +/- 1 °C

pH:

control 7.2 to 8.3, test concentration replicates 7.4 to 8.5

Dissolved oxygen:

7.8 - 9.3 mg O2 /l

Nominal and measured concentrations:

dilution levels 1/16; 1/8; 1/4; 1/2; and 1/1 (nominal) of the saturated solution, average measured concentrations (0h to 96 h) by TOC analysis: 0.46 mg/L, 0,87 mg/L, 1,79 mg/L, 3.68 mg/L and 7.13 mg/L respectively.

Details on test conditions:

TEST SYSTEM
- Test vessel: 20 L tank
- Material, size, headspace, fill volume: not specified
- Aeration: continous aeration
- Renewal rate of test solution: daily
- No. of organisms per vessel: 10 fish/vessel
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 10 fish with an average of 0.37 g
- Source/preparation of dilution water: drinking water
- Photoperiod: 16 h light - 8h darkness

EFFECT PARAMETERS MEASURED: mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: factor 2 for dilution

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

1.8 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

3.2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

7.1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The highest concentration without mortality after 96h is 1.8 mg/L (=LC0). The lowest test substance concentration with 100% mortality after 96 h is 7.1 mg/L (=LC100). All concentrations are based on the product. A 95% confidence interval is not specified as the the results has been determined by graphical analysis.

Validity criteria fulfilled:

not specified

Remarks:

The mortality of the control is <10%. Constant conditions were maintained. However, a stabillity control of the substance concentration was not be performed as the test organisms increased the carbon content. All concentration are based on the product.

Conclusions:

In a semi static test according to EC 92/69 part C1, the acute toxicity of 1,2,4-Trivinylcyclohexane on Brachydanio rerio has been examined. The LC50 after 96 h is 3.2 mg/L. The LC0 is 1.8 mg/L (highest concentration without observed mortality after 96 h). The LC100 is 7.1 mg/L (lowest concentration with 100% observed mortality after 96 h).

Description of key information

An acute toxicity test with the zebrafish revealed a LC50 of 0.65 mg/L after 96 h.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.65 mg/L

Additional information

The toxicity of the test substance has been examined according to OECD TG 203 and EU method C.1 with substance specific analysis. According to OECD guidance document No. 23 for substances that are difficult to test, a saturated solution has been tested with a nominal loading of 100 µg/L prepared by a slow-stirring procedure. First as a threshold approach, a semi-static limit test with the undiluted saturated solution has been performed. Since the saturated solution with a nominal loading of 100 µL/L has been found to cause mortality after 72 h, a concentration response test has been performed and further dilution levels of the saturated solution have been tested for 96 hours (semi-static test design). The dilution levels 6.25 %, 12.5 %, 25 %, and 50 % of the saturated solution were tested and no mortality has been observed in the second test.

The concentration of the test substance has been monitored by analytical measurement via GC-MS and the measured concentration at the end of each exposure interval (24 h) have been in the range of 52 % to 82 % of the initial concentration. The concentrations based on the geometric mean were 844 µg/L (100 % of the saturated solution), 504 (50 %), 241 µg/L (25 %), 126 µg/L (12.5%), and 62.2 µg/L (6.25 %). All effect levels were given based on the geometric mean measured concentrations of the test substance.

For evaluation and reporting the result of both tests have been regarded to cover the full concentration-response range. The LC50 after 96 hours has been determined to 652 µg/L (Conf. interval 504 – 844 µg/L). The LC0 and LC100 after 96 hours were 504 and 844 µg/L, respectively. The validity criteria of the study have been met. However, the effect level and the two highest tested concentration levels were above the determined water solubility of the test substance. Furthermore, the dilution levels have been prepared by dilution of the saturated solution instead of preparation of separated concentration levels. By this procedure that presence or enrichment of impurities due to the study design cannot be excluded.

 

In a supporting study, the acute toxicity of 1,2,4-Trivinylcyclohexane on fish has also been examined according to the guideline EEC 92/69 C1 and OECD203 (1984), but without substance specific analysis. In the semi static test with the zebrafish, a LC50 of 3.2 mg/L after 96 h has been observed. The highest test substance concentration without any mortality has been 1.8 mg/L (LC0). The analysis of the test substance concentration has been performed by TOC analysis and the results were based on the measured product concentrations.