Reaction product of 6H-dibenz[c,e][1,2]oxaphosphorin 6-oxide, itaconic acid and ethylene glycol

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:

2012-05-09 - 2013-01-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The GLP study was conducted according to an internationally accepted guideline. All study parameters are given in detail.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

The temperature in the main study was in the range of 20.2 - 22.7°C instead of 23 ± 2 °C.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

The temperature deviation in the main study was 2.5 °C instead of 2 °C (± 1 °C) as stated in the study plan.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

yes

Remarks:

The stock solutions prepared in acetone had a concentration between 1602 mg/L and 1608 mg/L instead of exactly 1600 mg/L as stated in the study plan.

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Vehicle:

yes

Details on test solutions:

During validation of the analytical method, the maximal solubility in Danio rerio test medium was determined as 1.6 mg/L. Because stability in test medium after 48 hours was only 81.4 % no saturated solution prepared Acetone was used as solvent to prepare the test solutions .at the limit of solubility directly before the start of the test and at medium renewal after 48 hours.
A stock solution containing 1600 ± 8mg/L in acetone was prepared. The concentration of the stock solution was 1000-fold higher than the concentration in the treatment (1.6 mg/L resp. the estimated maximal solubility in Danio test medium). For the solvent control, acetone in a concentration of 1 mL/L was used.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Danio rerio

Variety: Hamilton-Buchanan

Age: sexually immature young fish, length 2 ± 1 cm

Origin:

The original animals were obtained from Umweltbundesamt (UBA) and used for breeding and production of eggs. The offspring of the original animals is used in the test.

Husbandry:

Danio rerio is routinely used for toxicity tests.

Before being used for the test, the fish are kept for twelve days under test conditions. During this period, mortality must stay below 5 %.

Food:three times a day; warm-water fish food and daphnids

Medium renewal: twice a week

Photo period: 12/12 hours, using neon tubes

Temperature: 23 ± 2 °C

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

1.08 mmol/l

Test temperature:

23 ± 2 °C

pH:

7.8 - 8.1

Dissolved oxygen:

7.9 - 8.6 mg/l

Nominal and measured concentrations:

1.6 mg/l (limit of water solubility)

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.85 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

>= 0.85 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 1.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The analytical determination of the test item after 96 hours showed a concentration slightly above the limit of quantification in the solvent control. This may be caused by remains of the test item in the detection aperture The recovery after 48 hours before medium renewal was 38 % of the measured start concentration. The recovery after 96 hours was 29 % of the measured concentration after the 48 h medium renewal. The correlation between nominal and measured concentration was good (85 % after 0 hours and 99 % after 48 h in fresh prepared medium).

Sublethal observations / clinical signs:

Analytical Determinations Part1 Nominal Concentration in mg/L Measured Concentration in mg/L Measured Concentration in mg/L Geometric Mean in mg/L 0h 48h oM 48h nM 96h Solvent control <LOQ <LOQ <LOQ 0.16 - 1.6 1.36 0.51 1.58 0.46 0.85

oM old Medium nM new Medium

Analytical Determinations Part2

Nominal Concentration in mg/L	%Recovery after48hoM	%Recovery after96h	%Nominal concentration after 0h	%Nominal concentration after 48h nM
Solvent control	-	-	-	-
1.6	37.76	29.37	85.19	98.69

oM old Medium nM new Medium

Mortalities

Nom.Conc. in	Total of	dead fish
mg/L	fish used	24h		48h		72h		96h
		new	cum.	new	cum.	new	cum.	new	cum.
Solvent con­	7	0	0	0	0	1	1	0	1
trol
1.6mg/L	7	0	0	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The following results for the test item Ukanol FR 70 could be determined:
96h NOEC ≥ 0.85 mg/L (geometric mean)
96h LC50 > 0.85 mg/L (geometric mean)

96h NOEC ≥ 1.6 mg/L (nominal)
96h LC50 > 1.6 mg/L (nominal)

Executive summary:

The acute toxicity of Ukanol FR 70 against Danio rerio Hamilton Buchanan was determined following EU-Method C.1 resp. OECD Guideline 203.

The main study was performed as semi-static limit test using a concentration of 1.6 mg/L, representing the maximal solubility in test medium. The maximal solubility in test medium was determined during validation of the analytical method. Acetone was used as solvent to prepare the test solutions at the limit of solubility. Seven fish were exposed to the test item for 96 hours. After 48 hours the test medium was renewed. In the solvent control and in the treatment no significant mortality was observed. The pH and the oxygen value of the water were normal. During validation of the analytical method, the test item had shown insufficient stability. The recovery after four days had been 68.2 %. Therefore, the study was performed under semi-static conditions. The content of the test item in the test solutions was determined at the beginning and at the end of the test and after 48 h before and after medium renewal using HPLC. The correlation between nominal and measured concentration in the freshly prepared test solution was good (85 % after 0 hours and 99 % after 48 h). The recovery of measured start concentration was 38 % after 48 hours and 29 % after 96 hours. The unexpectedly poor recovery means that the characteristic of the test medium in presence of the animals is not identical with the medium without fish, used during validation of the analytical method. Without much doubt, the loss of test item was caused by ingestion by the fish or adsorption on to the animals or the excrements of the animals.

The following results for the test item Ukanol FR 70 could be determined:

96h NOEC ≥ 0.85 mg/L (geometric mean)

96h LC50 > 0.85 mg/L (geometric mean)

96h NOEC ≥ 1.6 mg/L (nominal)

96h LC50 > 1.6 mg/L (nominal)

For risk assessment purposes a NOEC value of 1.6 mg/l (nominal) will be used based on loss of the test substance in the test medium.

Description of key information

The following results for the test item Ukanol FR 70 could be determined:

96h NOEC ≥ 0.85 mg/L (geometric mean)

96h LC50 > 0.85 mg/L (geometric mean)

96h NOEC ≥ 1.6 mg/L (nominal)

96h LC50 > 1.6 mg/L (nominal)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.6 mg/L