Reaction mass of 1-[(1R*,6S*)-2,2,6-trimethylcyclohexyl]hexan-3-ol and 1-[(1S*,6S*)-2,2,6-trimethylcyclohexyl]hexan-3-ol

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:

key study

Study period:

04 - 08 Sep 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Staatliches Gewerbeaufsichtsamt Hildesheim, Hildesheim, Germany (03.01.2017)

Analytical monitoring:

yes

Remarks:

GC-MS

Details on sampling:

- Concentrations: Control and limit concentration of 1.50 mg/L (nominal)
- Sampling method: The limit concentration and the control were analytically verified in all fresh and aged test media.
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until sample preparation and at room temperature until the start of the analysis (on an autosampler), if necessary.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A saturated solution with a nominal loading of 1.50 mg/L test item was prepared by adding 1.67 µL/L test item (relative density considered) to dilution water and stirring slowly for 48 h at room temperature. The saturated solution was taken from the homogenous phase and was used for testing.
- Controls: Dilution water without test item subjected to the same test conditions.
- Evidence of undissolved material: No undissolved test item was observed during the test.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Length at study initiation: 2.69 cm (average)
- Weight at study initiation: 0.2 g (average)
- Holding: Fish were held at the test facility at 23 ± 2 °C and diffuse light (7 - 750 lux, natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80% of the air saturation value.

ACCLIMATION
- Acclimation period: At least 12 d
- Acclimation conditions: Same as test
- Feeding during acclimation: Fish were fed Sera Vipan (Sera GmbH, Heinsberg Germany) in 4% of the fish body weight, 3 times per week
- Health during acclimation: No mortality was observed, no disease treatment was necessary.

FEEDING DURING TEST
Fish were not fed during the test. Feeding was stopped 24 h before test start.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

58 mg/L CaCO3

Test temperature:

23.0 - 23.9 °C (control)
23.6 - 24.9 °C (treatment)

pH:

7.14 - 7.75 (control)
7.34 - 7.66 (treatment)

Dissolved oxygen:

74 - 98% oxygen saturation (control)
83 - 99% oxygen saturation (control)

Conductivity:

138 µS/cm

Nominal and measured concentrations:

Control, and 1.50 mg/L (nominal)
< LOQ , and 0.999 mg/L (geom. mean measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: 13 L glass aquaria (25.5 * 18 * 28 cm), filled with 8 L of dilution water (depth of water: 17 cm) and covered with a floating lid (e.g. aluminium foil).
- Aeration: None
- Renewal rate of test solution: 24 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 7
- No. of vessels per control (replicates): 7
- Biomass loading rate: 0.18 g fish/L test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water of local origin, filtered on activated charcoal and aerated for min. 24 h to remove chlorine.
- pH: 6.0 - 8.5
- Alkalinity: 0.6 mmol/L
- Acidity: 0.1 mmol/L
- Conductivity: 138 mS/cm
- Culture medium different from test medium: Dilution water same as holding water.
- Intervals of water quality measurement: The pH value, temperature and oxygen saturation were measured in all test vessels at the start of the test and at the end of the test and every 24 h and after renewal of the test media from fresh and old test media. Total hardness of the water was determined at the beginning in the control. Water temperature was recorded once per hour. Light intensity on the surface of the test aquaria was measured at the start of exposure.

OTHER TEST CONDITIONS
- Photoperiod: 16 h photoperiod daily
- Light intensity: 56 lux (mean value)

EFFECT PARAMETERS MEASURED:
- Mortality: After 3, 24, 48, 72, and 96 h
- Size and weight: After 96 h (control fish)

TEST CONCENTRATIONS
- Spacing factor for test concentrations: Threshold approach with one limit concentration of 1.50 mg/L (nominal, saturated solution)
- Range finding study : No range-finding study was conducted but a stability test was carried out with the saturated solution with a nominal loading rate of 1.50 mg/L to determine the optimal stirring time for maximum concentration.
- Results used to determine the conditions for the definitive study: No range-finding study was conducted. The threshold concentration is based on the toxicity data of an alga and a daphnid toxicity study.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.999 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: All fish were affected by sub-lethal effects. Quiescence was observed after 3 h. Missing escape reflex was recorded after 24 and 48 h. Slowed down and unregular breathing was recorded at 48, 72, and 96 h. Lethargy was recorded after 72 and 96 h.
- Mortality of control: 0%
- Other adverse effects control: None, normal behaviour throughout the test.
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No undissolved test item was observed.
- Effect concentrations exceeding solubility of substance in test medium: No

Sublethal observations / clinical signs:

VALIDITY CRITERIA

The study fulfilled the validity criteria set out by OECD guideline 203 (Table 1). Oxygen saturation was 74% (criterium: ≥ 60%), mortality in the control was 0% (criterium: max. 1/7 fish), and the pH in the control group deviated by 0.61 units (criterium: ≤ 1.0).

 

Table 1: Validity criteria for OECD 203.

Criterion from the guideline	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	Mortality in the control was 0% (0/7 fish)	Yes

 

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Conclusions:

The test resulted in an LC50 (96 h) of > 0.999 mg/L (geom. mean measured), indicating that the substance does not have lethal effects on zebrafish up to the threshold concentration of the saturated solution (OECD 203, D. rerio).

Executive summary:

There is one study, in which the short-term toxicity of test item to fish was assessed according to OECD 203 and GLP. In a semi-static limit test with a 24-h test medium renewal rate, D. rerio was exposed to one threshold concentration, consisting of a saturated solution with a nominal concentration of 1.50 mg/L, for 96 h. The concentration of the test item was analytically verified by GC-MS in fresh media (0, 24, 48, 72 h) and in aged media (24, 48, 72, 96 h).

The measured concentrations of the two main constituents of the test item at the end of each exposure interval ranged from 67 – 78% of the initially measured concentrations. The nominal threshold concentration of 1.50 mg/L corresponds to an overall geometric mean measured concentration of 0.999 mg test item/L. After 96 h, sub-lethal effects were observed and no mortality. Therefore, the LC50 (96 h) corresponds to > 0.999 mg test item/L (geom. mean measured concentration).

Description of key information

No effects up to saturation level (LC50 (96 h) > 0.999 mg/L, geom. mean measured, OECD 203, D. rerio)

Additional information

There is one study, in which the short-term toxicity of test item to fish was assessed according to OECD 203 and GLP. In a semi-static limit test with a 24-h test medium renewal rate, D. rerio was exposed to one threshold concentration, consisting of a saturated solution with a nominal concentration of 1.50 mg/L, for 96 h. The concentration of the test item was analytically verified by GC-MS in fresh media (0, 24, 48, and 72 h) and in aged media (24, 48, 72, and 96 h).

The measured concentrations of the two main constituents of the test item at the end of each exposure interval ranged from 67 – 78% of the initially measured concentrations. The nominal threshold concentration of 1.50 mg/L corresponds to an overall geometric mean measured concentration of 0.999 mg test item/L. After 96 h, sub-lethal effects were observed and no mortality. Therefore, the LC50 (96 h) corresponds to > 0.999 mg test item/L (geom. mean measured concentration).