1,6-Bis(methoxybenzoyloxy)hexane

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 January 2004 - 10 March 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study includes data generated according to generally valid and internationally accepted testing guidelines and performed according to GLP. The study was performed according to the following guidelines: - European Economic Community (EEC), EEC directive 92/69, Part C: Methods for the determination of ecotoxiciity, Publication No. L383, December 1992, C.2. "Acute Toxicity for Daphnia". - Organization for Economiic Co-operation and Development (OECD), OECD guidelines for Testing of Chemicals, guideline No. 202 Part I: "Daphnia sp., Acute Immobilisation Test", Adopted April 4, 1984. In addition, thenstudy procedures were based on the ISO International Standard 6341: "Water quality - Determination of the inhibition of the mobility of Daphnia magna Straus - Acute toxicity test, Third edition, 1996-04-01.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Remarks:

GLP Statement dates: 10 March 2004 and 12 March 2004

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
The concentrations at the starlt of the test were 0.07%-0.08% of the loaded concentration. The concentrations decreased by more than 20% during the test. At the end of the test, the concentrations were 21% and 27% of initial..

- Sampling method:
During the limit test duplicate samples were taken from-the test concentration and the blankcontrol.
Frequency At at t= 0 hr and t = 48 hr
Sample volume: 10 ml from the approximate centre of the test vessel

- Sample storage conditions before analysis:
Samples were transferred to Analytical Chemistry directly after sampling



Each sample (10 ml) was supplied in a 20 ml volumetric flask and filled up to the mark using
i;icetonitrile.

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The standard test procedures required generation of test solutions, which should contain completely dissolved test substance concentrations or stable and homogeneous mixtures or dispersions. The testing of concentrations that disturb the test system should be prevented (e.g. film of the test substance on the water surface).

The batch of Setafix X 11342 tested was a white/yellow crystal solid with a purity of >95% and not completely soluble in test medium at the concentrations tested.

A pre-test was performed to examine the solubility of the test substance in the test medium. A weighed amount of 10 mg was added to 1000 ml test medium. Magnetic stirring or ultra sonication did not accelerate 1the dissolving or dispersing of the test substance. Subsequently, acetone was used as a pre-solvent, but the substance did not dissolve in pure acetone at a concentration of 100 mg/ml.

In the limit test, a stock was prepared at 100 mg/I applying 72 hours of magnetic stirring. The resulting solution was clear and contained precipitate and a floating layer. After filtration (ca. 5 μm), the resulting test solution was clear and colourless.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: Crustacea, Cladocera (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions.
- Source: In-house laboratory culture with a known history
- Characteristics: Young daphnia with an age of < 24 hours were selected for the test, from parental daphnids of more than two weeks old.
- Reason for selection: This system has been selected as an internationally accepted invertebrate species.
- Validity of batch: Daphnids originated from a healthy stock, 2nd to 51th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.


Breeding:
- Start of each batch: With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
- Renewal of the cultures: After 7 days of cultivation half of the medium twice a week.
- Maximum age of the cultures: 4 weeks
- Medium: M7, as prescribed by Dr. Elendt-Schneider (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal
damage in Daphnia magna Straus. Protoplasma 154, 25-33).
- Temperature of medium: 18 - 22°C
- Feeding: Daily, a suspension of fresh water algae

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

At 24 hours and at 48 hours

Hardness:

250 mg/I expressed as CaCO3

Test temperature:

20.3 - 21.0°C

pH:

8.0 - 8.1

Dissolved oxygen:

8.6 - 8.7 mg/I

Salinity:

Not specified

Nominal and measured concentrations:

Nominal Concentration:
0 mg/I and 100 mg/I

Measured Concentration:
Initial measured concentration was 0.077 mg/I
During the exposure period the measured concentration had decreased to 0.019 mg/I

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 ml, all-glass
- Number of daphnia: 20
- Loading: 5 per vessel containing 80ml medium
- Feeding No feeding
- Introduction of daphnia: Within 40 minutes after preparation of the test solution
- Aeration: No aeration of the test solutions
- Test type: Static
- Test duration: 48 hours
- No. of organisms per vessel: 5 per vessel
- No. of vessels per concentration (replicates): quadruplicate
- No. of vessels per control (replicates): quadruplicate

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water purified by reverse osmosis (milli-RO, Millipore Corp., Bedford, Mass., USA)
- Metals: B, Fe, Mn, Li, Rb Sr and Mo
- Ca/Mg ratio: 2.5 : 1:
- Culture medium different from test medium: Culture medium is same as test medium (ISO-medium)

OTHER TEST CONDITIONS
- Adjustment of pH: No. pH was maintaiined within the limits prescribed by the guidelines.
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Effect parameters based on the average exposure concentration in a filtrate prepared at a loading rate of 100 mg/I are as follows:
Observed NOEC = < 0.038 (Concentration of Setafix X 11342 in mg/l)
24h-EC50 = < 0.038 (Concentration of Setafix X 11342 in mg/l)
48h-EC50 = < 0.038 (Concentration of Setafix X 11342 in mg/l)

TEST CONCENTRATIONS:
- Daphnia magna exposed to a filtrate prepared at a loading rate of 100 mg/I during the 48-hour test period.

- Analysis of the samples taken during the limit test showed that the mean measured concentration in the samples taken from the filtrate at the start of the test was 0.077 mg/I. During the exposure period the measured concentration had decreased to 0.019 mg/I (24 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/l (NOTOX Project 330918).

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr20 7, art. 4864, batch no. K28974764)

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 0.038 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: Immobilisation

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.038 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

other: Immobilisation

Dose descriptor:

NOEC

Effect conc.:

< 0.038 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: Immobilisation

Remarks on result:

other: Duration for NOEC not specified

Details on results:

- Measured concentrations:
Analysis of the samples taken during the limit test showed that the mean measured concentration in the samples taken from the filtrate at the start of the test was 0.077 mg/I. During the exposure period the measured concentration had decreased to 0.019 mg/I (24 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/l (NOTOX Project 330918).

Based on the analytical results, the calculated average exposure corwentration was 0.038 mg/I

- Effect parameters based on the average exposure concentration in a filtrate prepared at a loading rate of 100 mg/I are as follows:
Observed NOEC = < 0.038 (Concentration of Setafix X 11342 in mg/l)
24h-EC50 = < 0.038 (Concentration of Setafix X 11342 in mg/l)
48h-EC50 = < 0.038 (Concentration of Setafix X 11342 in mg/l)

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values:
No undissolved material was observed during the test

- Effect concentrations exceeding solubility of substance in test medium:
No EC50 could be calculated because the test substance proved to be non-toxic (EC50 > maximum soluble concentration).

Results with reference substance (positive control):

- Results with reference substance valid? Yes
The actual responses in this reference test with K2Cr20 7 are within the ranges of the expected responses at the different concentrations. Hence, the sensitivity of this batch of D. magna was in agreement with the historical data collected at NOTOX.

-The 24h-EC50 was 0.97 mg/I with a 95% confidence interval between 0.86 and 1.2 mg/I
- The 48h-EC50 was 0.66 mg/I with a 95% confidence interval between 0.57 and 0.83 mg/I

Validity criteria fulfilled:

yes

Remarks:

The study met the acceptability criteria prescribed by the protocol

Conclusions:

Under the conditions of the present study Setafix X 11342 did not induce more than 25% immobilisation of Daphnia magna exposed to a filtrate prepared at a loading rate of 100 mg/I during the 48-hour test period. The average exposure concentration was 0.038 mg/I, which was above water solubility. Hence, the 48h-EC50 for Daphnia magna exposed to Setafix X 11342 was above the water solubility limit of 8.8 μg/I.

Executive summary:

Acute Toxicity Study in Daphnia. ma.gna. with Setafix X 11342.

The study procedures described in this report were based on the ISO International Standard 6341, 1996. In addition, the procedures were designed to meet the test methods and validity criteria of the EEC directive 92/69, Part C.2, 1992 and the OECD guideline No. 202 Part I, 1984.

The batch of Setafix X 11342 tested was a white/yellow crystal solid with a purity of >95% and not completely soluble in test medium at the concentrations tested.

A limit test was performed exposing 20 daphnia in quadruplicate (5 per vessel) to a filtered solution prepared at a loading rate of 100 mg/l and a blank-control. The test solution was magnetically stirred for 72 hours, after which it was filtered over a paper filter (ca. 5 μm). The filtrate was clear and colourless. The total test period was 48 hours. Samples for analysis were taken at the start and the end of the test.

Analysis of the samples taken during the limit test showed that the mean measured concentration in the samples taken from the filtrate at the start of the test was 0.077 mg/l. During the exposure period the measured concentration had decreased to 0.019 mg/l (24 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/l (NOTOX Project 330918).

Based on the analytical results, the calculated average exposure corwentration was 0.038 mg/l

The study met the acceptability criteria prescribed by the protocol and was considered valid.

Under the conditions of the present study Setafix X 11342 did not induce more than 25% immobilisation of Daphnia magna exposed to a filtrate prepared at a loading rate of 100 mg/l during the 48-hour test period. The average exposure concentration was 0.038 mg/l, which was above water solubility. Hence, the 48h-EC50 for Daphnia magna exposed to Setafix X 11342 was above the water solubility limit of 8.8 μg/l

Description of key information

Under the conditions of the present study Setafix X 11342 did not induce more than 25% immobilisation of Daphnia magna exposed to a filtrate prepared at a loading rate of 100 mg/l during the 48-hour test period. The average exposure concentration was 0.038 mg/l, which was above water solubility. Hence, the 48h-EC50 for Daphnia magna exposed to Setafix X 11342 was above the water solubility limit of 8.8 μg/l

Additional information

Acute Toxicity Study in Daphnia. magna. with Setafix X 11342.

The study procedures described in this report were based on the ISO International Standard 6341, 1996. In addition, the procedures were designed to meet the test methods and validity criteria of the EEC directive 92/69, Part C.2, 1992 and the OECD guideline No. 202 Part I, 1984.

The batch of Setafix X 11342 t1ested was a white/yellow crystal solid with a purity of >95% and not completely soluble in test medium at the concentrations tested.

A limit test was performed exposing 20 daphnia in quadruplicate (5 per vessel) to a filtered solution prepared at a loading rate of 100 mg/l and a blank-control. The test solution was magnetically stirred for 72 hours, after which it was filtered over a paper filter (ca. 5 μm). The filtrate was clear and colourless. The total test period was 48 hours. Samples for analysis were taken at the start and the end of the test.

Analysis of the samples taken during the limit test showed that the mean measured concentration in the samples taken from the filtrate at the start of the test was 0.077 mg/l. During the exposure period the measured concentration had decreased to 0.019 mg/l (24 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/l (NOTOX Project 330918).

Based on the analytical results, the calculated average exposure concentration was 0.038 mg/l

The study met the acceptability criteria prescribed by the protocol and was considered valid.

Under the conditions of the present study Setafix X 11342 did not induce more than 25% immobilisation of Daphnia magna exposed to a filtrate prepared at a loading rate of 100 mg/l during the 48-hour test period. The average exposure concentration was 0.038 mg/l, which was above water solubility. Hence, the 48h-EC50 for Daphnia magna exposed to Setafix X 11342 was above the water solubility limit of 8.8 μg/l