Exo-1,7,7-trimethylbicyclo[2.2.1]hept-2-yl propionate

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

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: read-across from a guideline study

Justification for type of information:

The acute fish toxicity of Isobornyl propionate is based on read-across from Isobornyl acetate. The documentation is presented in the Aquatic Endpoint summary. The accompanying files are also attached there.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

8.3 mg/L

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

Effect value from the source chemical was recalculated based on Log Kow and molecular weight for the target chemical.

Validity criteria fulfilled:

yes

Remarks:

The information is used from read across. The read across rationale is documented according to Annex XI criteria.

Conclusions:

Tha acute fish toxicity for Isobornyl propionate is 8.3 mg/l using conversion from Isobornyl acetate results.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

8 Aug 2011 to 12 Aug 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Information is used for read across to Isobornyl propionate

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Version / remarks:

1996

Qualifier:

according to guideline

Guideline:

other: ASTM Standard E729-96: Standard Guide for Conducting Acute Toxicity Tests on Test Materials with Fishes, Macroinvertebrates and Amphibians

Version / remarks:

2007

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

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 three days. Samples of stock solutions were also taken at this time to confirm stock concentrations. 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. Two sets of samples were collected at 96 hours. One set was processed immediately for analysis, and the second set was stored refrigerated for possible future analysis. The samples were collected from mid-depth, acidified with two drops of phosphoric acid, placed in glass vials, and processed immediately for analysis.

Vehicle:

yes

Remarks:

0.1 mL/L dimethylformamide

Details on test solutions:

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 220 mg/mL. Four secondary stock solutions were prepared in DMF at nominal concentrations of 14, 28, 55 and 110 mg/mL by proportional dilution of the primary stock. The stock solutions were mixed by inversion, and ranged from clear and colorless to clear and light yellow in appearance. Stock solutions were stored refrigerated in glass amber bottles or glass graduated cylinders, and aliquots of each stock were placed in the syringe pumps every other day during the study. The five test substance stock solutions were injected into the diluter mixing chambers at a rate of 20.0 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. The concentration of DMF in the solvent control and all test item treatment groups was 0.1 mL/L.

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Source: Chesapeake Cultures, Inc., Hayes, VA 23072
- Age: All fish used in the test were juveniles from the same source and year class.
- Length at study termination in negative control: 3.1 - 4.2 cm (mean = 3.5 cm)
- Weight (wet weight) at study termination in negative control: 0.17 - 0.53 g (mean = 0.29 g)

ACCLIMATION
- Acclimation period: The fish were held for at least 14 days prior to the test.
- Acclimation conditions: same as test
- Feeding: Daily during the holding period, except during periods of fasting prior to testing, the fish were fed a commercially-prepared diet supplied by Sera North America, Montgomeryville, Pennsylvania, supplemented with brine shrimp nauplii (Artemia sp.) supplied by Brine Shrimp Direct, Ogden, Utah. The fish were not fed for at least two days prior to the test or during the test.
- Health during acclimation: During the 2-week period prior to the test the fish showed no signs of disease or stress.

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

132 mg/L as CaCO3 (at study initiation)

Test temperature:

21.5 - 22.0 °C

pH:

8.0 - 8.1

Dissolved oxygen:

5.8 - 8.0 (A dissolved oxygen concentration of 5.2 mg/L represents 60% saturation at 22ºC in freshwater)

Conductivity:

346 µS/cm (at study initiation)

Nominal and measured concentrations:

- Nominal test concentrations: 0 (control), 0 (solvent control), 1.4, 2.8, 5.5,11 and 22 mg/L
- Mean measured concentrations:

Details on test conditions:

TEST SYSTEM
- Test vessel: 25-L stainless steel aquaria filled with 15 L of test water. The depth of the test water in a representative chamber was 17.3 cm.
- Type of flow-through: A continuous-flow diluter was used to deliver each concentration of the test substance, a solvent control and a negative control (dilution water) to test chambers. Syringe pumps (Harvard Apparatus, Massachusetts) were used to deliver test substance stock solutions or solvent to impartially assigned mixing chambers where the stocks or solvent were mixed with dilution water prior to delivery to the test chambers. The flow of dilution water into each mixing chamber was controlled using rotameters and was adjusted to provide approximately 10 volume additions of test water in each test chamber per day. After mixing, the flow from each mixing chamber was split to deliver test water to two replicate test chambers. The syringe pumps used to deliver stock solutions or solvent to the mixing chambers, and the rotameters used to control the flow of dilution water to the mixing chambers were calibrated prior to the test. The proportion of the test water that was split into each replicate test chamber was checked prior to the test to ensure that flow rates varied by no more than ± 10% of the mean flow rate for the two replicates. Delivery of test solutions to the test chambers was initiated six days prior to the introduction of the test organisms to the test water in order to achieve equilibrium of the test substance. The general operation of the exposure system was checked visually at least once on the first and last days of the test and at least two times per day during the test.
- No. of organisms per vessel: 10
- No. of vessels per concentration: 2
- No. of vessels per control: 2
- No. of vessels per vehicle control: 2
- Biomass loading rate: 0.20 g fish/L/day test water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The water used for culturing and testing was 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 µm, 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 µm to remove fine particles and was passed through an ultraviolet (UV) sterilizer.
- Metals: Please see 'Any other information on materials and methods incl. tables' for an overview of the metals analysed in the test medium.
- Pesticides: Please see 'Any other information on materials and methods incl. tables' for an overview of the pesticides and organics analysed in the test medium.
- Chlorine: 3.5 mg/L
- Alkalinity: 184 mg/L as CaCO3 (at study initiation)
- Ca/mg ratio: 2.7
- Culture medium different from test medium: same as test

TEST MEDIUM MEASUREMENTS
- Temperature: The target test temperature during the study was 22 ± 1°C. Temperature was measured in each test chamber at the beginning and end of the test using a liquid-in-glass thermometer. Temperature also was monitored continuously in one negative control test chamber using a Fulscope ER/C Recorder, which was verified prior to test initiation with a liquid-in-glass thermometer.
- Dissolved oxygen and pH: Dissolved oxygen and pH were measured in one replicate test chamber of each treatment and control group at the beginning of the test, at approximately 24-hour intervals during the test, and at the end of the test, with measurements typically alternating between replicates in each group at each measurement interval. Dissolved oxygen was measured using a Thermo Orion Model 850Aplus dissolved oxygen meter, and measurements of pH were made using a Thermo Orion Model 525Aplus pH meter. When 100% mortality occurred in a test chamber, measurements of temperature, dissolved oxygen and pH were taken in that test chamber and then discontinued.
- Hardness, alkalinity, conductivity: Hardness, alkalinity and specific conductance in the dilution water were measured at the beginning of the test. Hardness and alkalinity measurements were made by titration based on procedures in Standard Methods for the Examination of Water and Wastewater. Specific conductance was measured using an Acorn Series Model CON6 conductivity-temperature meter.

OTHER TEST CONDITIONS
- Illumination: Ambient laboratory light was used to illuminate the test systems. Fluorescent light bulbs that emit wavelengths similar to natural sunlight.
- Light intensity: Light intensity at test initiation was 356 lux at the surface of the water of one representative test chamber. Light intensity was measured at the water surface of one representative test chamber at test initiation using a SPER Scientific Model 840006C light meter.
- Photoperiod: 16 hours of light and 8 hours of darkness, controlled by an automatic timer. A 30-minute transition period of low light intensity was provided when lights went on and off to avoid sudden changes in lighting.

EFFECT PARAMETERS MEASURED: mortality and sub-lethal effects
All organisms were observed periodically to determine the number of mortalities in each treatment group. The numbers of individuals exhibiting signs of toxicity or abnormal behavior also were evaluated. Observations were made approximately 2.5, 24, 48, 72 and 96 hours after test initiation.

TEST CONCENTRATIONS
- Results used to determine the conditions for the definitive study: Nominal test concentrations were selected based on the results of exploratory range finding toxicity data which indicated an LC50 value of >23 mg/L, and a functional solubility test which indicated a functional solubility of 21.2 mg/L in freshwater.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

9.9 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.I.: 7.5 - 13.0 mg/L

Remarks:

The LC50 was determined as geometric mean from the LC0 and LC100 value (mean meausured concentrations)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

3.3 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

MORTALITY
All fish in the negative and solvent control groups appeared normal throughout the test, with no mortalities. All fathead minnows in the 1.1 and 3.3 mg/L treatment groups also appeared normal throughout the test, with no mortalities or overt signs of toxicity observed. A single mortality was noted in the 1.6 mg/L treatment group at initial observations on Day 0. Based upon the timing of this mortality and lack of mortality at the 3.3 and 7.5 mg/L test concentrations, the mortality in the 1.6 mg/L treatment group was not considered to be related to treatment and was excluded from LC50 calculations. With the exception of the incidental mortality, all fathead minnows in the 1.6 mg/L treatment group appeared normal throughout the test, with no mortalities or overt signs of toxicity observed. Percent mortality in the 7.5 and 13 mg a.i./L treatment groups at test termination was 0 and 100%, respectively. A detailed overview of the mortality and sub-lethal effects is provided in 'Any other information on results incl. tables'.

SUB-LETHAL EFFECTS
Signs of toxicity observed among fish in the 7.5 and 13 mg/L treatment groups were surfacing and loss of equilibrium.

TEST SOLUTIONS
The test solutions in the test chambers appeared clear and colorless during the test, with no evidence of precipitation observed in any control or treatment solution.

Reported statistics and error estimates:

The mortality data were analyzed using the computer program 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, 48, 72 and 96-hour LC50 values and binominal probability was used to calculate the 95% confidence intervals. 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:

Table: Cumulative mortality and observations

Mean Measured Concentration (mg/L)	Rep.	No. Exposed	~2.5 Hours		24 Hours		48 Hours		72 Hours		96 Hours		Cumulative Percent Mortality
			No dead(1)	Observations(2)	No dead(1)	Observations(2)	No dead(1)	Observations(2)	No dead(1)	Observations(2)	No dead(1)	Observations(2)
Negative Control	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0		0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
Solvent Control	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0		0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
1.1	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0		0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
1.6	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0		5
	B	10	1	9 AN	1	9 AN	1	9 AN	1	9 AN	1
3.3	A	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0		0
	B	10	0	10 AN	0	10 AN	0	10 AN	0	10 AN	0
7.5	A	10	0	10 AN	0	8 AN; 2 A	0	7 AN; 3 A	0	8 AN; 2 A	0	7 AN; 3 A	0
	B	10	0	9 AN; 1 A	0	8 AN; 2 A	0	6 AN; 2 A; 2 N	0	7 AN; 3 A	0	5 AN; 4 A; 1 N
13	A	10	6	4 N	10	--	10	--	10	--	10	--	100
	B	10	8	2 N	10	--	10	--	10	--	10	--

1) Cumulative number of dead fish.

2) Observations: AN = appear normal; A = surfacing; N = loss of equilibrium.

Validity criteria fulfilled:

yes

Remarks:

see 'Any other information on materials and methods incl tables'.

Conclusions:

The 96-h LC50 value is 9.9 mg/L in fathead minnow (Pimephales promelas).

Executive summary:

The short-term toxicity to freshwater fish was determined in a GLP-compliant guideline study according to OECD TG 203. In this study, groups of fathead minnow (P. promelas) were exposed to nominal test substance concentrations of 0 (control), 0 (solvent control), 1.4, 2.8, 5.5,11 and 22 mg/L for 96 hours under flow-through conditions (20 organisms per treatment). The test item solution was prepared from individual stock solutions of calculated a amount of the test item mixed into HPLC-grade dimethylformamide (DMF) which was used as solvent (0.1 mL/L). Analysis of the test media at 0, 48 and 96 hours showed geometric mean measured test concentrations of <LOQ (control), <LOQ (solvent control), 1.1, 1.6, 3.3, 7.5 and 13 mg/L. The measured test concentrations did not remain remain within ±20% of nominal concentrations throughout the test and hence the results are based on the geometric mean measured test concentrations. Incidences of mortality and clinical effects were recorded after ~2.5, 24, 48, 72 and 96 hours exposure. With the exception of the incidental mortality, all fathead minnows in the 1.6 mg/L treatment group appeared normal throughout the test, with no mortalities or overt signs of toxicity observed. Percent mortality in the 7.5 and 13 mg/L treatment groups at test termination was 0 and 100%, respectively. Signs of toxicity observed among fish in the 7.5 and 13 mg/L treatment groups were surfacing and loss of equilibrium. Based on these findings, the 96-h LC0 is determined at 7.5 mg/L and the 96-h LC100 at 13 mg/L. The 96-h LC50 was calculated as geometric mean of the LC0 and LC100 (mean measured concentrations) and determined to be 9.9 mg/L. In this study all OECD validity criteria were met.

Description of key information

Isobornyl propionate

The acute fish toxicity of the Isobornyl propionate is 8.3 mg/l, which is a converted value from Isobornyl acetate based on the difference in Log Kow and molecular weight ( Log EC in mmol target= Log ECmmol x log Kow source/Log Kow target)

Isobornyl acetate and acute fish toxicity

For Isobornyl acetate the acute fish toxicity is derived from Isobornyl acetate(CAS 125 -12 -2). Itsshort-term toxicity to freshwater fish was determined in a GLP-compliant guideline study according to OECD

TG 203. In this study, groups of fathead minnow (P. promelas) were exposed to nominal test substance concentrations of 0 (control), 0 (solvent control), 1.4, 2.8, 5.5, 11 and 22 mg/L for 96 hours under flow-through conditions (20 organisms per treatment). The test item solution was prepared from individual stock solutions of calculated a amount of the test item mixed into HPLC-grade dimethylformamide (DMF) which was used as solvent (0.1 mL/L). Analysis of the test media at 0, 48 and 96 hours showed geometric mean measured test concentrations of <LOQ (control), <LOQ (solvent control), 1.1, 1.6, 3.3, 7.5 and 13 mg/L. The measuredtest concentrations did not remain remain within ±20% of nominal concentrations throughout the test and hence the results are based on the geometric mean measured test concentrations. Incidences of mortality and clinical effects were recorded after ~2.5, 24, 48, 72 and 96 hours exposure. With the exception of the incidental mortality, all fathead minnows in the 1.6 mg/L treatment group appeared normal throughout the test, with no mortalities or overt signs of toxicity observed. Percent mortality in the 7.5 and 13 mg/L treatment groups at test termination was 0 and 100%, respectively. Signs of toxicity observed among fish in the 7.5 and 13 mg/L treatment groups were surfacing and loss of equilibrium. The 96-h LC50 was calculated as geometric mean of the LC0 and LC100 (mean measured concentrations) to be 9.9 mg/L. In this study all OECD validity criteria were met.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

8.3 mg/L