Allyl phenoxyacetate

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:

11 - 15 Jan 2016

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

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

2008

GLP compliance:

yes (incl. QA statement)

Remarks:

Staatliches Gewerbeaufsichtsamt Hildesheim, Germany

Analytical monitoring:

yes

Details on sampling:

- Concentrations: test concentrations and the control were analytically verified from freshly prepared media after 0 and 72 h and from corresponding aged test media after 24 and 96 h
- Sample storage conditions before analysis: samples were stored at room temperature until the start of the analysis, if necessary. Prepared samples were stored in the autosampler at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: appropriate volumes of the test item were pipetted directly into the test vessels, filled with appropriate volumes of the dilution water, mixed with an ultraturrax (1 min, 12000 rpm) and covered with a floating lid
- Controls: dilution water without test item
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): No

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: fish used in the test were gained from a single brood stock of the laboratory (supplier: Umweltbundesamt, Berlin, Germany)
- Length at study initiation (mean): 2.29 cm
- Weight at study initiation (mean): 0.173 g

ACCLIMATION
- Method of holding: at 23 ± 2 °C, diffuse light (0.1-10 µmol photons * m^-2 * s^-1, natural photoperiod), water was changed at least once per week, dissolved oxygen concentration was more than 80% of the air saturation value
- Acclimation period: at least 12 days
- Type and amount of food: food: Sera Vipan; Sera GmbH, Heinsberg, Germany; amount: 4% of the fish body weight per feeding day. Test fish were not be fed 24 h before test start
- Feeding frequency: 3 times per week
- Health during acclimation (any mortality observed): No mortality and no disease treatments were administered throughout holding

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

85 mg CaCO3/L

Test temperature:

21.9 - 22.5 °C

pH:

6.51 - 6.74

Dissolved oxygen:

Oxygen saturation: 91 - 100%

Nominal and measured concentrations:

0 (control), 0.0625, 0.125, 0.25, 0.5 and 1 mg/L (nominal)
0 (control), 0.0384, 0.115, 0.154, 0.254 and 0.728 mg/L (geometric mean measured)

Details on test conditions:

TEST SYSTEM
- Test vessel: 64 L glass aquaria
- Material, size, headspace, fill volume: test aquaria were covered with a floating lid (i.e. with aluminium foil), fill volume was 40 L
- Aeration: no
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.03 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water of local origin (water was filtered on activated charcoal and aerated for at least 24 h to remove chlorine)
- Holding medium different from test medium: no
- Intervals of water quality measurement: pH, temperature and oxygen saturation were measured in all test vessels every 24 h from fresh and old test media. Total hardness of the water was determined at the beginning out of the control. Water temperature was recorded once per hour. Light intensity on the surface of the test aquaria was measured at test start.

OTHER TEST CONDITIONS
- Adjustment of pH: yes, to 6.50 ± 0.20 (repeated for each renewal day)
- Photoperiod: 8 h
- Light intensity: 1.79 to 2.35 µmol photons * m^-2 * s^-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : observations were made after 2, 24, 48, 72 and 96 h

TEST CONCENTRATIONS
- Range finding study: yes, semi static conditions, without headspace
- Test concentrations: 0 (control), 0.1, 1 and 10 mg/L
- Results used to determine the conditions for the definitive study: 100% mortality was observed after already 24 h at the two highest concentrations while no mortality took place after 96 h at the 0.1 mg/L concentration. Based on these results concentrations between 0 and 1000 µg/L were chosen for the definitive test.

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.133 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: based on the geometric mean of LC100/LC0

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

0.115 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

0.154 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: no

Sublethal observations / clinical signs:

Table 1: % mortality during exposure time in the definitive test.

Geometric mean measured concentration [mg/L]	Cumulative Mortality [%] after different exposure periods [h]
	2	24	48	72	96
0.728	0	100	100	100	100
0.254	0	0	100	100	100
0.154	0	0	57	100	100
0.115	0	0	0	0	0
0.0384	0	0	0	0	0
Control	0	0	0	0	0

The measured concentrations in the stability test were 94%, 46% and 28% of the nominal concentration after 0, 24 and 48 h, respectively.

Table 2: Measured concentrations and % of the nominal values of the test item.

Sampling date	0 h (Fresh medium)		24 h (Old medium)		72 h (Fresh medium)		96 h (Old medium)
Nominal concentration of the test item [mg/L]	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Geometric Mean Measured test item concentration* [mg/L]
1	0.780	78	0.679	68	-	-	-	-	0.728
0.5	0.286	57	0.226	45	-	-	-	-	0.254
0.25	0.141	56	0.169	68	-	-	-	-	0.154
0.125	0.0742	59	0.0557	45	0.211**	169	0.203**	162	0.115
0.0625	< LOQ**	-	< LOQ	-	0.0664	106	0.0525	84	0.0384
Control	< LOQ		< LOQ		< LOQ		< LOQ
* concentrations < LOQ were taken into account with 1/2 LOQ
** result not conclusive probably due to an application error but confirmed by repetition, thus used for calculation of the mean measured concentration of the test item

Validity criteria fulfilled:

yes

Conclusions:

The study (OECD 203, GLP) with the zebrafish Danio rerio resulted in a LC50 (96 h) of 0.133 mg test item/L (based on the geometric mean of LC100/LC0).

Executive summary:

The study was conducted according to OECD guideline 203, EU regulation No. 440/2008/ Method C.1 and GLP. The test organism, Danio rerio was exposed in a semi-static freshwater system to mean measured test item concentrations of 0 (control), 0.0384, 0.115, 0.154, 0.254 and 0.728 mg/L for 96 h. The definitive test was designed based on the findings of a range finding test with concentrations between 0.1 and 100 mg/L and an analytical test item stability test with 4 mg/L. The test concentrations were evaluated by HPLC-DAD analysis in freshly prepared media after 0 and 72 h and in the corresponding 24 h aged test media after 24 and 96 h. The measured concentrations vary between 45 and 169% of the nominal values and therefore all effect values base on geometric mean measured test item concentrations. Effects were observed in the three highest tested concentrations resulting in an LC50 (96 h) of 0.133 mg/L.

Description of key information

LC50 (96 h) = 0.133 mg/L (D. rerio, OECD 203) 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.133 mg/L

Additional information

One study is available, in which the short term toxicity of the substance to fish is tested. The study was conducted according to OECD guideline 203, EU regulation No. 440/2008/ Method C.1 and GLP. The test organism, Danio rerio was exposed in a semi-static freshwater system to mean measured test item concentrations of 0 (control), 0.0384, 0.115, 0.154, 0.254 and 0.728 mg/L for 96 h. The definitive test was designed based on the findings of a range finding test with concentrations between 0.1 and 100 mg/L and an analytical test item stability test with 4 mg/L. The test concentrations were evaluated by HPLC-DAD analysis in freshly prepared media after 0 and 72 h and in the corresponding 24 h aged test media after 24 and 96 h. The measured concentrations vary between 45 and 169% of the nominal values and therefore all effect values base on geometric mean measured test item concentrations. Effects were observed in the three highest tested concentrations resulting in an LC50 (96 h) of 0.133 mg/L.