Ethyl 2-(dimethylamino)-1-phenylcyclohex-3-ene-1-carboxylate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study in compliance with GLP-criteria.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Ethyl trans-(±)-2-(dimethylamino)-1-phenylcyclohex-3-ene-1-carboxylate hydrochloride

EC Number:

248-226-0

EC Name:

Ethyl trans-(±)-2-(dimethylamino)-1-phenylcyclohex-3-ene-1-carboxylate hydrochloride

Cas Number:

27107-79-5

IUPAC Name:

27107-79-5

Details on test material:

- Substance name: Trans-Tilidin-HCl-Hemihydrate
- Physical state/appearance: Solid (powder)/white
- Homogeneity: Homogeneous
- Storage conditions: At room temperature, under nitrogen atmosphere, in the dark and closed tightly

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples for analytical determination of the concentration were taken within the first hour after the start of exposure and at the end of the exposure after approx. 96 hours. Water samples were taken from the middle of the test vessel using a glass pipette and were transported to the analytical Iaboratory in glass ampoules, which were rinsed with test solution before they were filled. The transport to the Analytical Laboratory was done on the day of sampling.

Test solutions

Vehicle:

no

Details on test solutions:

The test concentrations were prepared separately for each test vessel. The test substance was weighed into a beaker. The defined volume of test water was filled into the test vessel. An aliquot from the test water in the test vessel was taken and mixed with the test substance in the beaker using an ultra turrax. The mixture was poured into the test vessel and the whole test batch was homogenized again with an ultra turrax. No solvents or dispersants were used tor the preparation of the test concentrations. All test batches were present as clear solutions over the exposure period according to visual inspection.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST SECIES
- Common name: Zebrafish
- Animal supplier: Zierfisch-Center Klöckner, Ludwigshafen, Germany
- Body weight (wet weight): 0.53 g (0.37 - 0.76 g)
- Body length (from the top of the snout to the end of the caudal fin): 3.9 cm (3.5 - 4.3 cm)
- Age of the animais: Approx. 4 months
- Hatching date: Jun. 24, 2002
- Arrival at the testing facility: Aug. 28, 2002
- All animals used for the study were derived from the same batch of fish.
- All test animals were observed for their health state during insertion into the test vessels. No signs for sickness, injuries or abnormalities were observed.

ACCLIMATION:
- Duration of acclimatization to testing conditions including test water and light regime: 14 days
- Photoperiod: 16 hours light, 8 hours dark
- Water quality: Non chlorinated charcoal filtered tap water (Frankenthal, Germany)
- Diet: Commercial fish diet "Tetramin", supplier Tetra-Werke, Melle, Germany, ad libitum, additionally generally on workdays live brine shrimp (artemia).
- Withdrawal of feed: During the last day before start of exposure
- Medical treatment: None during acclimatization
- Mortality during the last week before start of exposure: 0 %

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

250 mg/L (as CaCO3)

Test temperature:

22-23 °C

pH:

7.8-8.3

Dissolved oxygen:

7.3-8.5 mg/L

Nominal and measured concentrations:

- Nominal concentrations: 0 (control), 5, 10, 22, 50 and 100 mg/L (based on range finding study).
- Measured concentrations: n.d., 4.3, 8.6, 18.7, 42.4 and 92.8 mg/L (mean value of analytical concentrations at t=oh and t=96h)

Details on test conditions:

TEST CONDITIONS:
- Test vessels: Glass aquaria (30 cm long, 22 cm wide, 24 cm high)
- Volume of water: 10 L
- Animals/test vessel: 10
- Loading (g fish/L water): 0.5
- Test vessel/concentration: 1
- Photoperiod: 16 hours light, 8 hours darkness
- Aeration: Slight
- Feeding: None

INSERTION OF ANIMALS:
- The test organisms were introduced into the test vessels according to a randomization plan prepared by the testing laboratory using a program of the Iaboratory data evaluation group of the testing facility.

OBSERVATIONS:
- Symptoms and lethality: The fish were observed for survival and toxic signs (changes in appearance, swimming behaviour and behaviour in comparison to the control group) within 1 hour after start of exposure and 4, 24, 48, 72 and 96 hours after start of exposure. Fish were considered dead if there was no visible movement and no reaction after touching. Dead fish were removed from the test vessels.
- Water parameter: Temperature, oxygen content and pH-value were measured in each test vessel within 1 hour after start of exposure and after 24, 48, 72 and 96 hours.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

18.7 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

22 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No symptoms were observed up to a concentration of 10 mg/L nominal. At higher concentrations apathy and swimming at the bottom were recorded.

Reported statistics and error estimates:

Finney, D.J., Probit Analysis; Cambr. Univ. Press, 3rd ed., 1971 (certain aspects of this method have been modified)

Any other information on results incl. tables

Sublethal observations / clinical signs:

Cumulative mortality

Measured concentration (mg/L)	No. of fish at start	No. of fish dead after 96h	Mortality (%)
0	10	0	0
4.31	10	0	0
8.56	10	0	0
18.73	10	5	50
42.40	10	10	100
92.80	10	10	100