Octahydro-4,7-methano-1H-inden-5-yl acetate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03/06/2010 - 09/07/2010

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study according to international guideline (OECD Guidleine 203) under GLP. Klimisch 2 is derived because of presence of oily slick in test concentrations being indicative for the test substance not fully soluble at the tested concentration and which may have resulted in a more worst case LC50 value.

Justification for type of information:

This information is used to derive the acute / chronic ratio for Cyclaprop, which is then applied to Cyclacet Dihydro (see for further details the Endpoint summary).

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 2.4, 5.3, 12, 25 and 56 mg/l (nominal)
- Sampling method: Samples were collected from one test chamber of each treatment and control group three days prior to the start of the test after conditioning the diluter for approximately 20 hours. Water samples also were collected from alternating replicate test chambers in each treatment and control group at the beginning of the test and at 48 and 96 hours (± 1 hour) to measure concentrations of the test substance. The samples were collected from mid-depth, placed in glass vials, and processed immediately for analysis.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Individual stock solutions were prepared for each of the five concentrations tested. A primary stock solution was prepared by mixing a calculated amount of test substance into HPLC-grade dimethylformamide (DMF) at a nominal concentration of 560 mg/mL. Four secondary stock solutions were prepared in DMF at nominal concentrations of 24, 53, 120 and 250 mg/mL by proportional dilution of the primary stock. The stock solutions were mixed by inversion, and appeared clear and colorless. Stock solutions were stored refrigerated in 250 or 500-mL glass amber bottles or a 250-mL glass volumetric flask, and aliquots of each stock were placed in the syringe every two days during the study.The five test substance stock solutions were injected into the diluter mixing chambers at a rate of 20 µL/minute where they were mixed with dilution water delivered at a rate of 200 mL/minute to achieve the desired test concentrations. The negative control received dilution water only. The solvent control was prepared by delivering HPLC-grade DMF to the mixing chamber for the solvent control.
- Controls: Yes, blanks and solvent controls
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Dimethylformamide (DMF)
- Concentration of vehicle in test medium (stock solution and final test solution(s) including control(s)): 0.1 ml/ml
- Evidence of undissolved material (e.g. precipitate, surface film, etc): At test initiation, an oily-slick was noted on the surface of the solutions in the mixing chambers of the 12, 25 and 56 mg/L test concentrations and on the surface of the solution in the 56 mg/L treatment group test chamber. At test termination, an oily-slick was noted on the surface of the solutions in the mixing chambers and on the bottom of the test chambers of the 12, 25 and 56 mg/L treatment groups.

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: fATHEAD MINNOW
- Source: Chesapeake Cultures, Inc., Hayes, Virginia, USA
- Age at study initiation (mean and range, SD): Juvenile
- Length (control at end of study): 2.6 - 3.3 cm
- Weight (control at end of study): 0.11 - 0,24 grams
- Feeding during test: no feeding during test

ACCLIMATION
- Acclimation period: 14 days
- Acclimation conditions: same as test
- Type and amount of food: a commercially-prepared diet supplied by Zeigler Brothers, Inc., Gardners, Pennsylvania, supplemented with brine shrimp nauplii (Artemia sp.) provided by Brine Shrimp Direct, Ogden, Utah.
- Feeding frequency: daily, no feeding at least 2 days prior to the start of the test.
- Condition: mortality <1% and no soigns of stress

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

144 mg/l CaCO3

Test temperature:

21.9 - 22.0ºC

pH:

8.2 - 8.3

Dissolved oxygen:

7.3 - 8.5 mg/l

Nominal and measured concentrations:

Nominal: 2.4, 5.3, 12, 25 and 56 mg/l
Measured (mean): 1.7, 4.2, 8.0, 12 and 18 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: test chamber
- Material, size, headspace, fill volume: teflon-lined, stainless steel aquaria of 25l filled with 15 l of test water
- Renewal rate of test solution (frequency/flow rate): 10 volume additions of test water/day
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 2
- Biomass loading rate: 0.13 g fish/l

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: freshwater obtained from a well approximately 40 meters deep located on the Wildlife International, Ltd. site. The well water was passed through a sand filter to remove particles greater than approximately 25 m, and pumped into a 37,800 L storage tank where the water was aerated with spray nozzles. Prior to use, the water was filtered to 0.45 m to remove fine particles.
- Total organic carbon:
- Metals: Below LOQ
- Pesticides: Below LOQ
- Alkalinity: 180 mg/l (CaCO3)
- Conductivity: 373 uS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: Temperature at start and end of the experiment, pH and dissolved oxygen were measured daily

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16 hours light, 8 hours darkness with a 30 minute transition period
- Light intensity: 378 lux

EFFECT PARAMETERS MEASURED: mortality was observed 5, 24, 48, 72 and 96 hours after the start of the experiment

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

6.7 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95%CL: 4.2 - 8.0 mg/l

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

15 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95%CL: 12 - 18 mg/l

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95%CL: 4.2 - 12 mg/l

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

6.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95%CL: 4.2 - 12 mg/l

Details on results:

- Behavioural abnormalities: At the end of the experiment, fish in the 8.0 mg/l treatment group showed signs of lethargy and surfacing
- Mortality of control: 0%
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No
- Effect concentrations exceeding solubility of substance in test medium: No

Results with reference substance (positive control):

Not relevant

Reported statistics and error estimates:

The mortality data were analyzed using the computer program of C. E. Stephan (5). The program was designed to calculate the LC50 value and the 95% confidence interval by probit analysis, the moving average method, and binomial probability with nonlinear interpolation. In this study, nonlinear interpolation was used to calculate the 24, 72 and 96-hour LC50 values and binominal probability was used to calculate the 95% confidence intervals. Binomial probability was used to calculate the LC50 value and the 95% confidence limits at the 48-hour interval. Due to the method used to calculate the 96-hour LC50 value, the slope of the dose response curve could not be calculated. The no-mortality concentration and NOEC were determined by visual interpretation of the mortality and observation data.

Sublethal observations / clinical signs:

Measured concentrations of Cyclaprop in test solution samples

Nominal test concentration (mg/l)	Sample number (558A-105-)	Sampling time (days)	Measured concentration^1(mg/l)	Percentage of nominal^2	Mean measured concentration (mg/l)	Mean percent of nominal
Negative Control	1 8 15	0 2 4	<LOQ <LOQ <LOQ	- - -	-	-
Solvent Control	2 9 16	0 2 4	<LOQ <LOQ <LOQ	- - -	-	-
2.4	3 10 17	0 2 4	1.74 1.70 1.74	72.6 71.0 72.5	1.7	72
5.3	4 11 18	0 2 4	4.23 4.00 4.28	79.8 75.4 80.7	4.2	79
12	5 12 19	0 2 4	9.49 7.27 7.18	79.1 60.6 59.8	8.0	67
25	6 13	0 2	12.3 11.6	49.0 46.3	12	48
56	7 14	0 2	18.1 17.3	32.3 30.9	18	32

 

¹The limit of quantification (LOQ) was 0.506 mg/l, calculated as the product of the concentration of the lowest calibration standard (0.500 mg a.i./L) and the dilution factor of the matrix blank samples (1.00), corrected for the test substance purity (98.6%).

²Results were generated using Excel 2000 in the full precision mode. Manual calculations may differ slightly.

Cumulative mortality and observations

Mean measured concentration (mg/)	Rep.	No. exposed	~5 Hours		24 Hours		48 Hours
			No. dead^1	Observations^2	No. dead^1	Observations^2	No. dead^1	Observations^2
Negative control	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0 0	10 AN 10 AN
Solvent control	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0 0	10 AN 10 AN
1.7	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0 0	10 AN 10 AN
4.2	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0 0	10 AN 10 AN
8.0	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	6 4	2 AN; 2 C 3 AN; 2 A; 1 C
12	A B	10 10	0 0	4 AN; 6 C 5 AN; 5 C	0 0	5 AN; 5 C 5 AN; 5 C	10 10	- -
18	A B	10 10	3 3	2 A; 3 C; 1 N; 1 R 3 C; 1 N; 3 R	10 10	- -	10 10	- -

 

Mean measured concentration (mg/)	Rep.	No. exposed	72 Hours		96 Hours		Cumulative percent mortality
			No. dead^1	Observations^2	No. dead^1	Observations^2
Negative control	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0
Solvent control	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0
1.7	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0
4.2	A B	10 10	0 0	10 AN 10 AN	0 0	10 AN 10 AN	0
8.0	A B	10 10	9 5	1 A 2 A; 2 C; 1 N	9 6	1 A 4 C	75
12	A B	10 10	10 10	- -	10 10	- -	100
18	A B	10 10	10 10	- -	10 10	- -	100

¹Cumulative number of dead fish

²Observations: AN = appear normal; A = surfacing; C = lethargic; N = loss of equilibrium; R = lying on the bottom of the chamber The concentration-response curve at 96h is included as an illustration below.

Validity criteria fulfilled:

yes

Remarks:

Mortality in controls <10%, oxygen concentration above 60% of saturation, results based on measured concentrations

Conclusions:

The acute toxicity (96h-LC50) of Cyclaprop towards Pimephales promelas is 6.7 mg/l.

Executive summary:

The acute toxicity of Cyclaprop towards Pimephales promelas was investigated according to OECD guideline 203 under GLP. Fish were exposed to nominal concentrations of 2.4, 5.3, 12, 25 and 56 mg/l under flow-through conditions and observed for 96 hours. Based on mean measured concentrations, the 96h-LC50 was found to be 6.7 mg/l.

The test concentrations where oily-slick was observed coincide with the concentrations at which mortality was found. Physical hindrance may therefore have contributed to the effects found. It is however, difficult to make a distinction betwen physical and systemic effects and therefore as a worst case approach the mortality is assumed to result from systemic toxicity.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

13 September 1993 to 17 September 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to EU method C.1 in compliance with GLP, without deviations that influence the quality of the results.

Justification for type of information:

The information of Cyclacet is used for read across to Cyclacet Dihydro (see for further details the Endpoint summary).

Reason / purpose for cross-reference:

read-across: supporting information

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 3, 6, 12, 24, 48 mg/l
- Sampling method: The test material concentration was measured in each test chamber at test initiation (before the addition of the fish) and after 24 hours.

Vehicle:

no

Details on test solutions:

The concentrations of the stock solutions were 96 or 150 mg/L. Appropriate aliquots were added directly to each test chamber to achieve the nominal concentrations 3, 6, 12, 24 and 48 mg/L.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Strain: Hamilton Buchanan
Source: West-Aquarium (Bad Lauterberg)
Age: 26 February 1993 (birth date), supply date of test organisms: 14 May 1993
Body length: 2.5 -3.5 cm
Diet: no medical pretreatment
Pre-test mortality of test organisms: < 2%
Water: synthetic water based on ISO, hardness: 13.2° dH

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

13.2 °dH

Test temperature:

21.6 - 23.3 °C

pH:

7.2 - 8.1

Dissolved oxygen:

7.6 - 8.7 mg/L

Nominal and measured concentrations:

The concentration of the test substance was measured two times during the test (test initiation and 24 hrs later). Maximal deviation from nominal concentration was - 20%.
Nominal vs Measured concentrations of test material at test initiation [mg/L]
3.0 vs 2.7
6.0 vs 4.5
12.0 vs 11.0
24.0 vs 22.5
48.0 vs 37.9

Details on test conditions:

TEST SYSTEM
- Test vessel: 6 different aquariums/test chambers of 300 x 135 x 200 mm (flow -through)
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): 6 full exchanges per day
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: synthetic freshwater (according to ISO)

OTHER TEST CONDITIONS
- Photoperiod: 14 hours light, 10 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Mortality after 2, 24, 48, 72, 96 hours

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

10.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

behaviour

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

23.1 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

15.8 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL 10.9 - 23.1 mg/l. Derived from reported data

Details on results:

LC0/LC100: 15.8 mg/L (geometric mean). This value can be used as the LC50.
Mortality of control: 0

Reported statistics and error estimates:

The LC 0 and LC 100 values were estimated based on visual inspection and using a standard operation procedure (SOP 4.002) which is not further described in the study report.

Sublethal observations / clinical signs:

No toxic effects were observed in the control.

No mortality in control and concentrations 3, 6, 12 mg/l. 100% mortality after exposure during 24 h in 24 and 48 mg/l.

Validity criteria fulfilled:

yes

Remarks:

Exposure concentrations less than 80% but results based on measured data, control mortality < 10%

Conclusions:

An LC50 value of 15.8 mg/L was determined for the mortality of the test substance for Brachydanio rerio. The 95%-confidence levels were 10.9 and 23.1 mg/l.

Executive summary:

The acute toxicity of Cyclacet towards Brachydanio rerio was investigated according to EU Method C.1 in compliance with GLP. Fish were exposed to nominal concentrations of 3, 6, 12, 24 and 48 mg/l under flow-through conditions and observed for 96 hours. No mortality was observed in the control and concentrations 3, 6 and 12 mg/l, whereas 100% mortality was reported after 24 h in 24 and 48 mg/l exposure. The LC50 is derived from the geometric mean of the LC0 and the LC100 value of (10.9 mg/l and 23.1 mg/l): 15.8 mg/l based on mean measured concentrations.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The information is based on read across.

Justification for type of information:

The full read across justification is presented in the Endpoint summary, the accompanying file is also attached there.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

15.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Validity criteria fulfilled:

yes

Executive summary:

This summary includes the information from other species. The full read across is presented in the Endpoint summary

Cyclacet Dihydro has the same acute aquatic toxicity as Cyclacet and has the same acute /chronic ratio as derived for the Cycla-esters. Cyclacet Dihydro is much alike this group of esters, which have a tricyclodecenyl fused ring and short alkyl chain esters attached to it. The ethyl- and propyl are called Cyclacet and Cyclaprop, respectively. The acute aquatic toxicity will be derived from Cyclacet. The chronic aquatic toxicity will be derived using the acute /chronic ratio derived from Cyclaprop.

Structural similarities and differences:The Cyclacet Dihydro and Cyclacetboth have a tricyclodec- fused ring structure with an acetic ester attached to the bridged hexyl-ring. Cyclaprop has a propyl ester attached to this fused-ring structure. The difference with Cyclacet is the missing of the double bond in the pentyl-ring (as presented in the name of Cyclacet Dihydro). The difference with Cyclaprop is the missing of the double bond in the pentyl ring and the attached ester is a propyl one instead of an acetic one. This means that both the backbone and the functional groups are very similar.

Bioavailability:Cyclacet Dihydro and Cyclacet have verysimilar bioavailability based on the similarity in chemical structure and physico-chemical properties. The molecular weight of both substances is 194 versus 192. They are both liquids, have very similar measured vapour pressures (2.2 and 2.1 Pa) and water solubilities (89 and 186 mg/l, they also have the same calculated log Kow. The measured log Kow of Cyclacet Dihydro is slightly higher (4.5) compared to Cyclacet (3.9) but is anticipated to the experimental variability. Overall, the bioavailability and the aquatic toxicity profile of the target and the source substances will be similar.

Mode of Action and the prediction of the chronic information:Cyclacet Dihydro has the same mode of action as the other Cycla-esters. Their similar backbone and their similar functional group, being an ester, will present a similar mode of action. This same mode of action has resulted in an incremental decrease of acute LC and EC50 values for fish and Daphnia based on the increased alkyl chain from Cyclacet onwards (see data matrix).

Acute aquatic toxicity: For the prediction of acute aquatic toxicity for Cyclacet Dihydro the experimentally data from Cyclacet are used.

Chronic aquatic toxicity: For prediction of the chronic toxicity of Cyclacet Dihydro for algae and micro-organisms the result from Cyclacet will be used.

For predicting the chronic toxicity of Daphnia and fish the Acute / Chronic ratio of Cyclaprop will be used. This reflects the toxicity best because a) experimental data are used; b) it reflects the difference in toxicity of the shorter and longer alkyl chains; c) the approach can be applied to all Cycla-esters and; d) the approach can be used for all species.

Fish: The key information is the experimental Cyclaprop A/C ratio of 8.4 for fish, which results in a Cyclacet Dihydro value of 1.9 ( 15.8 (mg/l) / 8.4 (A/C ratio). This value will be used for the risk assessment.

Daphnia: The key information is the experimental A/C ratio for Daphnia of 14 for Cyclaprop, which results in a Cyclacet Dihydro EC10/NOEC of 1.8 mg/l (25 (mg/l) / 14). This 1.8 mg/l will be used for the risk assessment.

Uncertainty of the predictions: For acute aquatic toxicity there is a high certainty for the read across between from Cyclacet Dihydro and Cyclacet in view of their similaritiesconsidering the structure, the physico-chemical properties and the mode of action.The A/C ratios for the different trophic levels for Cyclacet Dihydro based on Cyclaprop have a high certainty because of close similarity between these two substances considering the structure, the physico-chemical properties and the mode of action. Therefore the alkyl chain length will not have an impact on the A/C ratio and thus the A/C ratio of Cyclaprop can be used to present the chronic toxicity of Cyclacet.

Description of key information

Acute toxicity to fish, LC50 is 15.8 mg/l, based on read across from Cyclacet.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

15.8 mg/L