Propanoic acid, 2-hydroxy-2-methyl-, 2-propen-1-yl ester

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Report date:

1999

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Principles of method if other than guideline:

US EPA FIFRA-Guidline: Number of replcants, of test organisms, continuous monitoring of temperature, and equilibrium time, concentrations are 50% of the next higher Concentrations (factor between concentrations equals 2).

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

prop-2-en-1-yl 2-hydroxy-2-methylpropanoate

EC Number:

700-908-3

Cas Number:

19444-21-4

Molecular formula:

C7H12O3

IUPAC Name:

prop-2-en-1-yl 2-hydroxy-2-methylpropanoate

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Sample storage conditions before analysis: Room Temperature

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The stock solution was prepared the following way:
0.587 g test item were mixed with and made up to 1000 ml with dilution water.
Calculated amounts of the stock solution to produce the desired test concentrations were given into the water. After the equilibration time of 4.4 hours, the fish were transferred into the aquaria (beginning of exposure). No additional test solvents were used in this study since CA 2215 A is soluable in water at the levels tested.

Test organisms

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow Trout.
- Source: Commercial supplier - P. Holer / CH-4314 Zeiningen / Switzerland.
- Length at study initiation : (mean) 46mm (range 40 - 58mm) based on 7 fish of the control.
- Weight at study initiation: (mean) 0.77g (range 0.50 - 1.48g) based on 7 fish of the control.

ACCLIMATION
- Acclimation period: Fish were gradually acclimated to the test conditions, and were held for 60 days prior to testing in water of similarqality used in the test.

- Feeding frequency: Fish were not fed 24 hours prior to the start of the test
- Health during acclimation : Total mortality did not exceed 3% during the 7 day period prior to the start of the test.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test conditions

Hardness:

Total hardness 160 mg CaCO3/L.

Test temperature:

The temperature was maintained constant at 12.8 - 13.4°C.

pH:

The pH was determined to be 8.0 - 8.4

Dissolved oxygen:

The dissolved oxygen content was between 88 - 100% of the saturation value by gentle aeration during exposure.

Nominal and measured concentrations:

The nominal concentrations were 0.58, 1.0, 1.8, 3.2 and 5.8 g/L. The measured concentrations of CA 2215 A were 0.520, 0.916, 1.77, 3.38 and 5.90 at the start and 0.481, 0.863, 1.59, 3.00 and 5.57 at the end of the test, respectively.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria of 20 liters filled with 15 liters of water (35/22/26 cm; depth of water: 20cm)
- Aeration: The water was gently aerated durng exposure (oil free air)
- No. of organisms per vessel: 7 per concentration and control. 7 per aquarium.
- No. of vessels per concentration (replicates): One test chamber per concentration
- No. of vessels per control (replicates): One test chamber
- Biomass loading rate: Fish biomass to solution ratio was 0.36g/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dechlorinated tap water of the town of Basel (carbon filtered to eliminate chlorine andparticulate material).
- Total organic carbon: 0.4 - 0.9mg/L
- Particulate matter: Dilution water was filtered through activated carbon prior to it's use, thus retaining suspended solids

EFFECT PARAMETERS MEASURED: At 2, 24, 48, 72, 96 hours, observations of mortality and sublethal symptoms, such as abnormal behavioural activity and stress were made (swimming bhaviour, loss of equilibrium, respiratory function, exophthalmus, igmentation and other observations). Dead fish, if any, were removed from the test solutions , at least at the above mentioned intervals. At test termination , the fish in the control were retained for measurements of weight and length.

OTHER TEST CONDITIONS
- Photoperiod: 16 hours of light and 8 hours od dark with a 30 minutes transition period.
- Light intensity: Fluorescent light

TEST CONCENTRATIONS

- Range finding study
Prior to initiating the definitive test, a static range finding test was performed at concentrations 0.1, 1.0, 10, 100 mg/L.
Based on the results of this range finding test, the following nominal concentrations were selected for the definitive test.
0.58, 1.0, 1.8, 3.2, 5.8 mg/LCA 2215 A A/L (nominal)

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 2.97 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % CL

Effect concentrations 2

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

ca. 3.15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % CL

Effect concentrations 3

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

ca. 3.46 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % CL

Effect concentrations 4

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

ca. 5.71 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95 % CL

Details on results:

Lethal effects: after 96 h exposure, mortality occured at concentrations of 3.2 and 5.8 mg/L with 86 and 100% mortality, respectively. A decrease of the LC50 value from 5.71 to 2.97 mg/L was observed drng the period 24 to 96 h exposure. The LC0 (96 h) was 1.8 mg/L and the LC100 (96 h) 5.8 mg/L. The mortalityin the blank control was 0%.

Sublethal effects were observed at concentrations ≥ 3.2 mg/L, such as change in pigmentation, loss of equilibrium, change in the swimming behaviour and in the respiratory function, hence, the highest concentration with no sublethal and lethal effects was 1.8 mg/L.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Table 1: Mortalities

Initial number of fish: 7 per test concentration

Conc. Nominal	Cumulative Mortality Number of dead fish
(mg/L)	2-4 h	24 h	48 h	72 h	96 h	% 96 h
Blank 0.58 1.0 1.8 3.2 5.8	0 0 0 0 0 0	0 0 0 0 0 4	0 0 0 0 1 7	0 0 0 0 4 7	0 0 0 0 6 7	0 0 0 0 86 100

 

Table 2: Sublethal Symptoms observed

Conc. nominal	Swimming behaviour					Loss of equilibrium					Respiratory function					Exophthalmus					Pigmentation
(mg/L)	2-4	24	48	72	96	2-4	24	48	72	96	2-4	24	48	72	96	2-4	24	48	72	96	2-4	24	48	72	96
Blank 0.58 1.0 1.8 3.2 5.8	0 0 0 0 0 0	0 0 0 0 0 2	0 0 0 0 1	0 0 0 0 2	0 0 0 0 2	0 0 0 0 0 0	0 0 0 0 0 2	0 0 0 0 1	0 0 0 0 2	0 0 0 0 2	0 0 0 0 0 0	0 0 0 0 0 2	0 0 0 0 0	0 0 0 0 2	0 0 0 0 2	0 0 0 0 0 0	0 0 0 0 0 0	0 0 0 0 0	0 0 0 0 0	0 0 0 0 0	0 0 0 0 0 0	0 0 0 0 0 1	0 0 0 0 1	0 0 0 0 1	0 0 0 0 1

 

Explanation:  0: no symptoms, 1: light symptoms, 2: moderate symptoms, 3: severe symptoms

Table 3: Measurement of Temperature, pH and Oxygen Saturation

Conc. nominal (mg/L)	0 h			24 h			48 h			72 h			96 h
	T	pH	O2	T	pH	O2	T	pH	O2	T	pH	O2	T	pH	O2
	°C		%	°C		%	°C		%	°C		%	°C		%
Blank 0.58 1.0 1.8 3.2 5.8	13.0 12.8 12.8 12.8 12.9 12.9	8.0 8.2 8.2 8.2 8.3 8.3	93 93 93 93 93 93	13.3 13.2 13.2 13.2 13.1 13.2	8.2 8.3 8.3 8.3 8.3 8.3	96 96 99 95 97 100	13.3 13.2 13.2 13.2 13.1 13.1	8.2 8.3 8.3 8.3 8.3 8.4	97 97 99 98 97 98	13.4 13.2 13.2 13.2 13.1	8.3 8.3 8.3 8.3 8.3	94 94 99 99 98	13.3 13.3 13.2 13.2 13.2	8.1 8.2 8.3 8.3 8.3	97 97 97 97 88

 

T: Temperature

Table 4: Analytically determined Concentrations of the test item

Conc. nominal (mg/L)	Measured Concentrations of CA 2215 A
	Initial (0 h)		End (96 h)		Mean		Standard Deviation	% of Mean
	(mg/L)	%	(mg/L)	%	(mg/L)	%	±	Initial (0h)	End (96h)
Blank 0.58 1.0 1.8 3.2 5.8	< 0.2 0.520 0.916 1.77 3.38 5.90	90 92 98 106 102	< 0.2 0.481 0.863 1.59 3.00 5.57	83 86 88 94 96	< 0.2 0.501 0.890 1.680 3.190 5.735	86 89 93 100 99	0.028 0.037 0.127 0.269 0.233	104 103 105 106 103	96 97 95 94 97
Mean Min Max		97 90 106		89 83 96		93 86 100		104 103 106	96 94 97

 

Values are correct for recovery rate.

*The specimens were taken after 48 h since all fish had died after 48 h exposure.

Applicant's summary and conclusion

Conclusions:

The LC50 (96h) of CA 2215 A was determined to be 2.97 mg/L based on nominal concentration concentrations.
The highest concentration with no sublethal and lethal effects was 1.8 mg CA 2215 A/L.
Based on similarity of the measured concentrations of CA 2215 A at the start and end of the exposure, the test item was stable in the test solutions over the period of the test under static conditions.

Executive summary:

Report:Acute Toxicity Test ofCA2215A(Intermediate ofCGA276854) to Rainbow Trout (Oncorhynchus mykiss) Under Static Conditions

Guidelines: OECD-Guideline No.: 203; 92/69/EECC. l; US EPA FIFRA-Guideline No. 72-1

 Deviations to Guidelines: US EPA FIFRA-Guideline:   Number of replicates, of test organisms, continuous monitoring of temperature, and equilibrium time, concentrations are 50% of the next hgher concentrations (factor between concentrations equals 2). 

GLP: Confirmed GLP- Laboratory

Materials & Methods:

 

Test Item:	Identification Code:	CA 2215 A
	Batch No.:	P.608005
	Purity:	99.0%
	Solubility (in water):	200 mg/L in water (20°C)
	Appearance:	Colourless liquid
	Stability:	July, 2000

 

 

Static pre-tests showed that the LC50 (96h) was between 0.1 and 100 mg/L. Therefore, the following nominal concentrations were chosen for the main test: 0.58, 1.0, 1.8, 3.2 and 5.8 test item/L. 

After the preparation of the test concentrations and an equilibrium time of 4.4 hours, 7 fish were randomly placed in each of the 20 L glass aquaria containing 15 L of test solution (beginning of exposure). Fish were exposed to CA 2215 A for 96 hours and were observed for adverse effects at 2, 24, 48, 72 and 96 hours. 

 

Findings:

The temperature was maintained constant at 12.8 – 13.4°C and the dissolved oxygen contents between 88 – 100 % of the saturation value by gentle aeration during exposure. The pH was determined to be 8.0 – 8.4 and the total hardness 160 mg/L. 

The test item appeared homogeneously distributed in the test vessels at all test concentrations over the period of the test. No precipitation was observed in the testing solutions throughout the duration of the test. The nominal concentrations were 0.58, 1.0, 1.8, 3.2 and 5.8 mg test item/L. The measured concentrations of CA 2215 A at the beginning and at the end of the exposure period ranged from 90 - 106% and 83 – 96% of the nominal concentrations respectively. 

The following calculations and values are based on nominal concentrations. 

 

Exposure period	LC50	95% confidence	Slope	Goodness of fit
(h)	(mg/L)	(mg/L)	(mg/L)	X^2	df	p
96 72 48 24	2.97 3.15 3.46 5.71	none none none none	14.2 12.6 13.7 12.1	0.00 0.00 0.00 0.00	3 3 3 3	1.00 1.00 1.00 1.00

 

 

After 96 hours exposure, mortality occurred at concentrations of 3.2 and 5.8 mg/L with 86 and 100% mortality, respectively. 

A decrease of the LC50-value from 5.71 to 2.97 mg/L was observed during the period 24 to 96 h exposure. The LC 0 (96h) was 1.8 mg/L and the LC 100 (96h) 5.8 mg/L. 

 

Sublethal effects were observed at concentrations ≥ 3.2 mg/L, such as change in pigmentation, loss of equilibrium, change in the swimming behaviour and in the respiratory function, hence, the highest concentration with no sublethal and lethal effects was 1.8 mg/L.