2-hydroxyethyl [(1R,2S,5R)-2-isopropyl-5-methyl-cyclohexyl] carbonate

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:

1994-04-18 to 1994-04-22

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

GLP study according to OECD 203 (1992): effect concentration based on nominal concentrations, although the amount of test item in old media is <80 % of nominal values; recalculation was done

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

(1992)

GLP compliance:

yes (incl. QA statement)

Remarks:

signed 1994-03-16

Analytical monitoring:

yes

Details on sampling:

- Sampling method: Samples were taken of fresh and the corresponding 24-h old media over the whole test period (96 hours).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Direct dispersion in water
- Controls: yes, one control

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Length at study initiation at test start: 4.2 cm (SD = 0.2 cm)
- Weight at study initiation at test start: 0.89 g (SD = 0.18 g)

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): yes
- Type and amount of food during acclimation: commercial trout pellets
- Feeding frequency during acclimation: daily
- Health during acclimation (any mortality observed): mortality <1 % prior to test start

FEEDING DURING TEST
None.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

approximately 100 mg/L as CaCO3

Test temperature:

14 °C

pH:

7.3 - 7.6

Dissolved oxygen:

9.9 - 10.2 mg O2/L

Nominal and measured concentrations:

Nominal concentrations: control, 1.8, 3.2, 5.6, 10, 18 mg/L
Measured concentrations (recalculation of mean measured): control, 1.39, 2.61, 4.68, 8.34, 15.8 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass aquaria holding 20 L test media
- Aeration: via narrow bore glass tubes
- Renewal rate of test solution: yes, daily
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.45 g bodyweight/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Photoperiod: 16h light: 8h dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: approx. 1.8
- Range finding study
- Test concentrations: 1.0 to 100 mg/L with 9 concentrations)
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

11.5 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: geometric mean of LC0/LC100 based on recalculation of mean measured concentrations

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

8.3 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Recalculation of arithmetic mean based on geometric mean of measured concentrations of each renewal period

Duration:

24 h

Dose descriptor:

LC100

Effect conc.:

15.8 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Recalculation of geometric mean of measured concentrations of the first renewal period only (all fish were dead after 24 h)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

2.6 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

behaviour

Remarks on result:

other: Recalculation of arithmetic mean based on geometric mean of measured concentrations of each renewal period

Details on results:

- Increased pigmentation at 4.68 mg/L (mean measured)

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

Analysis of the mortality data by the moving average method of Thompson (1947).

Sublethal observations / clinical signs:

Cumulative Mortality

Mean Measured Concentration* (mg/L)	Cumulative mortality (initial population: 10)
	3h	6h	24h	48h	72h	96h
Control	0	0	0	0	0	0
1.39	0	0	0	0	0	0
2.61	0	0	0	0	0	0
4.68	0	0	0	0	0	0
8.34	0	0	0	0	0	0
15.8	0	0	10	10	10	10

 *Recalculation

Behavioural Observations

Mean Measured Concentration* (mg/L)	Abnormality	Time (hours)
		3	6	24	48	72	96
Control	No abnormalities detected
1.39	No abnormalities detected
2.61	No abnormalities detected
4.68	Increased pigmentation						3/10
8.34	Swimming at surface. Swimming at bottom. Increased pigmentation		3/10	3/10	3/10       5/10	5/10     10/10	5/10     10/10
15.8	Swimming at surface. Swimming at bottom. Increased pigmentation	7/10   3/10   10/10	7/10   3/10   10/10	A/D

*Recalculation

A/D = All fish dead

Analytical results for the test item

Samples	Nominal concentration of test item (mg/L)	Concentration found of test item found (mg/L)	Concentration of test item expressed as % of nominal
0 hours (fresh media)	Control 1.8 3.2 5.6 10 18	<L.O.D. 1.65 2.93 5.85 8.91 15.6	- 91.6 91.7 104.4 89.1 86.6
24 hours (old media)	Control 1.8 3.2 5.6 10 18	<L.O.D. 1.27 2.47 4.63 7.49 16.0	- 70.6 77.1 82.7 74.9 89.0
24 hours (fresh media)	Control 1.8 3.2 5.6 10	<L.O.D. 1.64 3.10 4.88 9.43	- 90.9 97.0 87.1 94.3
48 hours (old media)	Control 1.8 3.2 5.6 10	<L.O.D. 1.25 2.34 4.42 8.30	- 69.7 73.0 78.9 83.0
48 hours (fresh media)	Control 1.8 3.2 5.6 10	<L.O.D. 1.61 3.02 5.23 9.03	- 89.5 94.4 93.4 90.3
72 hours (old media)	Control 1.8 3.2 5.6 10	<L.O.D. 1.09 2.12 3.49 *	- 60.3 66.3 62.3 *
72 hours (fresh media)	Control 1.8 3.2 5.6 10	<L.O.D. 1.80 3.06 5.02 9.12	- 100.2 95.7 89.6 91.2
96 hours (old media)	Control 1.8 3.2 5.6 10	<L.O.D. 1.02 2.09 4.24 7.01	- 56.6 65.2 75.8 70.1

*Sample lost due to breakage

L.O.D. = Limit of Detection

Recalculation of mean measured test item concentration (arithmetic mean) based on geometricmean of the measured test item concentrations (fresh media - old media) for each renewal period.

Nominal test concentration	1.8 mg/L	geometric mean	3.2 mg/L	geometric mean	5.6 mg/L	geometric mean	10 mg/L	geometric mean	18 mg/L	geometric mean
0 h (fresh media)	1.65		2.93		5.85		8.91		15.6
24 h (old media)	1.27	1.45	2.47	2.69	4.63	5.20	7.49	8.17	16.0	15.8
24 h (fresh media)	1.64		3.10		4.88		9.43		all fish dead
48 h (old media)	1.25	1.43	2.34	2.69	4.42	4.64	8.30	8.85	all fish dead
48 h (fresh media)	1.61		3.02		5.23		9.03		all fish dead
72 h (old media)	1.09	1.32	2.12	2.53	3.49	4.27	*	-	all fish dead
72 h (fresh media)	1.80		3.06		5.02		9.12		all fish dead
96 h (old media)	1.02	1.35	2.09	2.53	4.24	4.61	7.01	8.0	all fish dead
geometric mean (24 h)										15.8
arithmetic mean (96 h)		1.39		2.61		4.68		8.34

*Sample lost due to breakage

Validity criteria fulfilled:

yes

Conclusions:

In this study a lethal effect on Oncorhynchus mykiss was observed at a mean measured concentration of 15.8 mg test item/L after 24 hours.
The 96-hour LC50 was determined to be 11.5 mg/L (mean measured), with a corresponding LC0 (96 h) and LC100 (24 h) of 8.3 mg/L and 15.8 mg/L, respectively.

Executive summary:

The acute toxicity of the test item to fish (Oncorhynchus mykiss) was determined under semi-static conditions with daily renewal of the test media over a period of 96 hours according to OECD 203 (1992). Based on result of the range-finding study, 5 nominal concentrations ranging from 1.8 to 18 mg/L were tested. Effect concentrations (LC50, LC0, LC100) were recalculated based on calculation of mean measured test item concentrations. Increased pigmentation was observed at 4.68 mg/L (mean measured) and 100 % mortality was already observed after the first test medium renewal period (0-24 hours) at 15.8 mg/L (mean measured). The 96-hour LC50 was determined to be 11.5 mg/L based on the calculated geometric mean measured concentrations attribute the toxic effect of LC0 (96 h) and LC100 (24 h).

Description of key information

The test item was found to be toxic to Oncorhychus mykiss.The 96-hour LC50 was determined to be 11.5 mg/L (mean measured), with a corresponding LC0 (96 h) and LC100 (24 h) of 8.3 mg/L and 15.8 mg/L, respectively.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

11.5 mg/L

Additional information

The acute toxicity of the test item to fish (Oncorhynchus mykiss) was determined under semi-static conditions with daily renewal of the test media over a period of 96 hours according to OECD 203 (1992). Based on result of the range-finding study, 5 nominal concentrations ranging from 1.8 to 18 mg/L were tested. Effect concentrations (LC50, LC0, LC100) were recalculated based on calculation of mean measured test item concentrations. Increased pigmentation was observed at 4.68 mg/L (mean measured) and 100 % mortality was already observed after the first test medium renewal period (0-24 hours) at 15.8 mg/L (mean measured). The 96-hour LC50 was determined to be 11.5 mg/L based on the calculated geometric mean measured concentrations attribute the toxic effect of LC0 (96 h) and LC100 (24 h).