3-phenylpropan-1-ol

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:

from 2016-02-16 to 2016-08-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: The limit concentration was diluted with mobile phase, prior to analysis. The control was analysed without dilution.
- Sampling method: The limit concentration and the control were analytically verified the test item via HPLC-DAD from freshly prepared media at the start and the end of the exposure. Additionally, the test concentration and the control were analytically verified via analysis of the dissolved organic carbon (DOC, according to DIN EN 1484) from freshly prepared media after 0 and from corresponding aged test media after 96 hours.
- Preparation of fortified samples: The fortified samples of the 1 x LOQ level and the blank samples were analysed without dilution. The 10 x LOQ level was diluted factor 5 with mobile phase, prior to analysis.
- Sample storage conditions before analysis: All samples were stored at 6 ± 2 °C until the start of the analysis. Prepared samples were stored in the autosampler at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: An appropriate amount of the test item was weighed and transferred to an aquaria filled with an appropriate volume of the dilution water. The test media was mixed with an ultraturrax (1 minutes, 17000 rpm).
- Controls: Blank control performed with the test solution without test substance.
- Evidence of undissolved material: No

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Source: All fish used in the test was gained at the test facility from a single brood stock (supplier: Umweltbundesamt, Schichauweg 58, D-12307 Berlin)
- Length: Average body length at the test start: 2.46 cm
- Weight: Average body weight at the test start: 0.184 g

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions: Holding was performed at the test facility at 23 ± 2 °C and diffuse light (0.1 - 10 pmol photons • nv2 - s-\ natural photoperiod). The water was changed at least once per week. The dissolved oxygen concentration was more than 80 % of the air saturation value.
- Type and amount of food during acclimation: Sera Vipan; Sera GmbH, 52518 Heinsberg, Germany. The amount of food was 4 % of the fish body weight per feeding day.
- Feeding frequency during acclimation: Food was provided 3 times per week. The test fish were not be fed 24 h before test start.
- Health during acclimation: The mortality was determined to be < 5 % within the last 7 days before the start of the exposure. No mortality was observed during this time. No disease treatments were administered throughout holding and testing.

FEEDING DURING TEST
No feeding was performed during the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Remarks on exposure duration:

None

Post exposure observation period:

Not performed

Hardness:

64 mg/L as CaCO3

Test temperature:

22.1 ± 0.2 °C

pH:

6.0 - 8.5

Dissolved oxygen:

Not less than 60 % of air saturation value.

Salinity:

Not applicable

Conductivity:

No data

Nominal and measured concentrations:

- Nominal concentration: 61 mg/L (limit test)
- Measured concentration: At 0 hours: 61.4 mg/L (101 % of nominal concentration) and at 96 hours 23.5 mg/L (39 % of nominal concentration)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria of 13 L were used, filled with approximately 10 L of dilution water (dimensions: 25.5/18/28 cm) and loosely covered with glass plates.
- Test volume: About 10 L per vessel.
- Aeration: Gentle aeration was provided in the test vessels.
- No. of organisms per vessel: 7 zebrafish were used each for the limit concentration and the control.
- No. of vessels per concentration (replicates): One replicate for the limit concentration
- No. of vessels per control (replicates): One replicate for the control
- Biomass loading rate: Fish density in the test vessels was 0.129 g fish per litre test solution.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Same as for holding.
- Culture medium different from test medium: No
- Intervals of water quality measurement: pH-value, temperature and oxygen saturation were measured all test vessels at the start of exposure and every 24 hours. Total hardness of the water was determined at the beginning out of the control. During the test the water temperature was recorded once per hour with a data logger.
The light intensity on the surface of the test aquaria was measured at test start.

OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: A daily 11.5 hours photoperiod was maintained throughout exposure, which corresponded to the natural daylight hours.
- Light intensity: 0.1 - 10 pmol photons • m-2 • s-1 (corresponding to approximately 7 - 750 Lux)

EFFECT PARAMETERS MEASURED: Mortality and visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.) after 2, 24, 48, 72 and 96 h.

TEST CONCENTRATIONS
- Range finding study: Not performed

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 61 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: The test substance was found to cause no effects to zebrafish after 96 hours at a nominal test item concentration of 61.0 mg/L.
- Observations on body length and weight: No
- Mortality of control: No
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: Not observed
- Effect concentrations exceeding solubility of substance in test medium: No
After 96 hours the LC50 was determined to be greater than 61 mg/L.

Results with reference substance (positive control):

No results with reference substance were performed.

Sublethal observations / clinical signs:

Table 1: Observations in the Definitive Test (n = 7)

 

Nominal test item concentration [mg/L]	Observations *)	Number
		2	24	48	72	96
61.0	(1)	0	0	0	0	0
Control	(1)	0	0	0	0	0

*) The numbers in brackets correspond to the following observations:

(1) = Normal behavior

 

 

Table 2: Cumulative Mortality [%] in the Definitive Test (n = 7)

 

Nominal test item concentration [mg/L]	Cumulative Mortality [%] after different exposure periods (hours)
	2	24	48	72	96
61	0	0	0	0	0
Control	0	0	0	0	0

 

  

Table 3: Measured Concentrations and Percent of the nominal Concentrations of the Test Item

 

Nominal concentration of the test item [mg/L]	Meas. conc. [mg/L]	[%]	Meas. conc. [mg/L]	[%]	Geometric Mean Measured test item concentration [mg/L]
61.0	61.0	101	23.5	39	38.0
Control	< LOQ		< LOQ

 

Meas. conc. = Measured concentration of the test item, dilution factor taken into account

% = Percent of nominal concentration of the test item

LOQ = Limit of Quantification (1 mg/L test item)

Table 4: Measured Organic Carbon Concentrations of the Control and the Test Item after 0 and 96 hours

(measured as DOC)

Nominal test item concentration [mg/L]	PHENYLPROPYL ALCOHOL
	Measured. conc.[mg C/L]	Measured conc.[mg C/L]
61.0	45.0	36.5
Control	< LOQ	2.62

LOQ = Limit of Quantification (2 mg C/L)

 

Validity criteria fulfilled:

yes

Conclusions:

In the acute toxicity of the test item to fish (zebrafish) according to OECD 203 (1992) the LC50 was determined to be > 61.0 mg/L (Threshold approach).

Executive summary:

The acute toxicity of the test item to fish (zebrafish) was determined according to the principles of the OECD-Guideline for Testing of Chemicals No. 203 (1992) and EU Council Regulation (EC) No. 440/2008/ Method C.1 (2008) from 2016-03-07 to 2016-05-13 with the definitive exposure phase from 2016-03-07 to 2016-03- 11 at the test facility.

The acute toxicity test under static conditions with one application at test start was conducted with the threshold concentration of 61 mg/L. Duration of the test was 96 hours. Seven test organisms were exposed to each test concentration and the control. Water quality parameters (i.e. temperature, pH-value and oxygen-saturation) measured at 0, 24, 48, 72 and 96 hours were determined to be within the acceptable limits.

The determination of the test item concentrations was carried out via HPLC-DAD. The evaluation was carried out from freshly prepared media after 0 and from the corresponding test media after 96 h. The measured concentration of the test item in freshly prepared media after 0 hours was 101 % of the nominal value. The measured concentration in the corresponding 96 h aged test media was 39 % of the nominal value. The concentration of the dissolved organic carbon (DOC), measured after 0 and 96 hours, showed a slight decrease at the end of the test. It is assumed that the decrease of the concentration of the test substance must have been caused by the formation of an unknown degradation product. In the chromatogram of the 96 hours old solution, an additional peak was observed at run time 1.00, indicating the formation of a more polar degradation product. Therefore, it is assumed that the test substance and its degradation product were still present, and all effect values are based on the nominal concentration of the test item.

Description of key information

In an acute toxicity test with zebrafish Danio rerio, the 96 h-LC50 of test item was determined to be greater than 61 mg/L (Threshold approach).

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

61 mg/L

Additional information

The acute toxicity of the test item to fish (zebrafish) was determined according to the principles of the OECD 203 (1992). The acute toxicity test under static conditions with one application at test start was conducted with the threshold concentration of 61 mg/L. Duration of the test was 96 hours. The determination of the test item concentrations was carried out via HPLC-DAD. The measured concentration of the test item in freshly prepared media after 0 hours was 101 % of the nominal value. The measured concentration in the corresponding 96 h aged test media was 39 % of the nominal value. The concentration of the dissolved organic carbon (DOC) at 0 h and 96 h were also measured and showed a slight decrease at the end of the test. Therefore, the 96h-LC50 was determined to be greater than 61 mg/L based on the nominal test item concentration.