1,6-Bis(methoxybenzoyloxy)hexane

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12-Jun-2010 to 16-Jun-2010

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 based on the OECD guidelines for Testing of Chemicals, guideline No. 203: "Fish Acute Toxicity Test", adopted 17 July, 1992

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Remarks:

GLP Statement date: 23 June 2010

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
At loading rate of 0 mg/L the concentrations of Z1077 in samples was < 0.1 mg/L
At a loading rate of 100 mg/L was as follows:

Time after Exposure (h) Concentration Analysed (mg/L)
0 1.10
24 1.05
48 0.967
72 0.934
96 0.891

Average exposure concentration was, therefore, calculated to be 0.994 mg/L

- Sampling method:
Samples were taken at 0 h, 24h, 48h, 72h and 95h from the test concentration and the blank-control. Sample of about 50 ml was taken each time
from each of the two vessels.

- Sample storage conditions before analysis:
Processed samples were stored in methanol solution at 4 oC for HPLC analysis

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

A weighed amount of 1000 mg test substance was added to each 10 L dilution. The resultant test solution was prepared by magnetically stirring of the dispersion for 8 hours, after which it was filtered over a paper filter.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebrafish
- Strain: Brachydanio rerio
- Source: obtained from Hangzhou Yangfan Tropic Fish Hatchery
- Length at study initiation (length definition, mean, range and SD): Average length of 3.04 cm
- Weight at study initiation (mean and range, SD): Average weight of 0.180 g
- - Method of breeding: The fish were domesticated in dechlorinated rust-free tap water of 23-25 oC. During the domestication, the fish were fed at least once daily until 24 hours before test.

- Feeding during test: Not fed

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): same as test
- Feeding frequency: daily
- Health during acclimation (any mortality observed): Total mortality of the population was record to be 5%

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

0 h, 24 h, 48 h, 72 h, and 96 hours after the exposure

Hardness:

60-80 as counted by CaC03

Test temperature:

23.1 - 24.3 oC

pH:

6.01 - 6.24

Dissolved oxygen:

6.79 - 6.3 mg/L

Salinity:

Not available

Nominal and measured concentrations:

Measured initial concentration was 1.10 mg/L. After 24 hours of exposure, the measured concentration was 1.05 mg/L. At the end of the 96h exposure period the measured concentration decreased to 0.891 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Test vessel: Glassware with diameter of 30 cm and height of 30 cm
- Type (delete if not applicable): closed
- No. of organisms per vessel: 10 fish per group, to be introduced into the diluent within 30 minutes after preparation
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 1
- Aeration: Diluent was not aerated during the test
- Photoperiod: 12h light vs 12h dark
- Light intensity: Fluorescent lamp, with light intensity of 600 - 800Lux
- Feeding: The fish were not fed during the total test period.

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
To prepare the diluent, a weighed amount of 1000 mg test substance was added to each 10 L dilution. After a stirring of 8 hours, the mixture was filtrated with qualitative filter paper to remove undissolved test substance particles.
The dilution was the same as that used for fish domestication


TEST CONCENTRATIONS
Chemical analysis showed that the measured initial concentration was 1.10 mg/L. After 24 hours of exposure, the measured concentration was 1.05 mg/L. At the end of the 96h exposure period the measured concentration decreased to 0.891 mg/L.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Dose descriptor:

NOEC

Effect conc.:

0.994 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Duration not specified

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 0.994 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 0.994 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 0.994 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

95 h

Dose descriptor:

LC50

Effect conc.:

> 0.994 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Poisoning symptoms:
-No observable symptoms were observed in both control and treatment group over the 96 h period

Mortality:
- No fish was found ded during the period of 96 hours

- Ther table shows the effect of Z1077 based on the average exposure concentration to the fish.

Effect of Z1077 to zebrafish in limit test
Parameter Concentration (mg/L)
Observed NOEC 0.994
(24 h) LC50 > 0.994
(48 h) LC50 > 0.994
(72 h) LC50 > 0.994
(95 h) LC50 > 0.994

N.B: Average concentration in a filtrate prepared at a loading rate of 100 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? YES
The response observed in fish originating from the present batch falls within this range.

Under the conditions of the present test potassium dichromate induced no lethal effects in fish at or below 223 mg/L. The 24h-LC50 was 316 mg/L. The range of the 24h-LC50 for fish is generally between 200 and 400 mg/L based on historical data of reference tests. The response observed in fish originating from the present batch falls within this range.

Validity criteria fulfilled:

yes

Conclusions:

Z1077 induced no visible effects in zebrafish exposed to a filtrate prepared at a loading rate of 100 mg/L. This suggested that LC50 of the test substance was above this concentration level. Due to its soluble limit, the average exposure concentration of Z1077 was measured to be 0.994 mg/L.

Executive summary:

The purpose of the study was to evaluate the test substance for its ability to generate acute toxic effects Z1077 in Brachydanio rerio during an exposure period of 96 hours and, if possible, to determine the LC₅₀ at all observation times. The study procedure was based on the OECD guidelines for Testing of Chemicals, guideline No. 203: "Fish Acute Toxicity Test", adopted 17 July, 1992.

The batch of Z1077 tested was a light yellow solid with a purity of >97% and not completely soluble in test medium at the concentrations tested.

The test was performed by exposing 10 fish to a filtered solution prepared at a loading rate of 100 mg/L and a blank-control. The test solution was prepared by magnetically stirring of the dispersion for 8 hours, after which it was filtered over a paper filter. The total test period was 96 hours. Samples for analysis were taken at the start, after 24, 48, 72, and 96 hours after the exposure.

Chemical analysis showed that the measured initial concentration was 1.10 mg/L. After 24 hours of exposure, the measured concentration was 1.05 mg/L. At the end of the 96h exposure period the measured concentration decreased to 0.891 mg/L.

Z1077 induced no visible effects in zebrafish exposed to a filtrate prepared at a loading rate of 100 mg/L. This suggested that LC₅₀ of the test substance were above this concentration level. Due to its soluble limit, the average exposure concentration of Z1077 was measure to be 0.994 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 January 2004 - 16 January 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 based on the ISO International Standard 7346-1: Static method, 1996 and also designed to meet the test methods and validity criteria of the EEC directive 92/69, Part C.1, 1992 and the OECD guideline No. 203, 1992.

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Remarks:

STATEMENT OF GLP COMPLIANCE Date: 12/03/2004

Analytical monitoring:

yes

Details on sampling:

- 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. After 24 hours of exposure, the measured conc~:mtration was 0.031 mg/I. During the remainder of the exposure
period the measured concentration decreased to 0.019 mg/I (on average 26 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/I (NOTOX Project 330918).

Based on the analytical results, the calculated average exposure concentration was 0.030 mg/I.

- Sampling method:
During the final test duplicate samples were taken from the test concentration and the blankcontrol.
Frequency: at t= Oh, t= 24 hand t= 96 h.

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

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION:
- Pre-liminary Treatment:
A pre-test was performed to e1xamine 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 the 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.

- Method:
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 (Schleicher and Schuell, ca. 5 μm) to remove the larger undissolved test substance particles, the resulting test solution was clear and colourless.

- Controls:
Test medium without test substance or other additives

- Chemical name of vehicle (organic solvent, emulsifier or dispersant):
Water

- Evidence of undissolved material (e.g. precipitate, surface film, etc):
Yes. After filtration (Schleicher and Schuell, ca. 5 μm) to remove the larger undissolved test substance particles, the resulting test solution was clear and colourless.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Strain: Cyprinus carpio
- Source: Zodiac, proefacc, "De Haar Vissen", LU. Wageningen, the Netherlands.
- Length at study initiation (length definition, mean, range and SD): 2.4 ±0.2 cm
- Weight at study initiation (mean and range, SD): 0.38 ± 0.09 g
- Feeding during test: No feeding from 48 hours prior to the test and during the total test period.

ACCLIMATION
- Acclimation period: 12 days
- Acclimation conditions (same as test or not): Yes
- Type of food: Trouvit.
- Feeding frequency: Daily
- Health during acclimation (any mortality observed): In the batch of fish used for the test, mortality during the seven days prior to the start of the test was less than 5%.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

At 4, 24, 48, 72 and 96 hours following the start of exposure.

Hardness:

250 mg CaC03

Test temperature:

20 - 24°C , constant within 2°C

pH:

6.0 - 8.5, constant within 1 unit

Dissolved oxygen:

oxygen> 60% of the air saturation value

Nominal and 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. After 24 hours of exposure, the measured concentration was 0.031 mg/I. During the remainder of the exposure period the measured concentration decreased to 0.019 mg/I (on average 26 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/I (NOTOX Project 330918).
Based on the analytical results, the calculated average exposure concentration was 0.030 mg/I

Details on test conditions:

TEST SYSTEM
- Test vessel: 10 litres, all-glass, containing 9.5 litres of test medium
- Type (delete if not applicable): closed
- Aeration: The test media were not aerated during the test.
- No. of organisms per vessel: 0.28 g fish/litre, i.e.7 fish per 9.5 litres of test medium
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water:
ISO-medium, formulated using Milli-Rowater (tapwater purified by reverse osmosis; Millipore Corp., Bedford, Mass., USA) with the following composition:
CaCl2.2H2O 293.8 mg/I
MgSO4.7H2O 123.3 mg/I
NaHCO3 64.8 mg/I
KCI 5.8 mg/I

- Culture medium different from test medium: No

- Intervals of water quality measurement:
pH, nitrate and nitrite concentration and ammonia concentration: once a week. Temperature: every day.

- Photoperiod: 16 hours photoperiod daily

DETERMINATION OF EFFECT PARAMETERS MEASURED:

Parameter Concentration Setafix X 11342 (mg/L)
Observed NOEC 0.030
24h LC50 > 0.030
48h LC50 > 0.030
72h LC50 > 0.030
96h LC50 > 0.030

The values above are average exposure concentrations in a filtrate prepared at a loading rate of 100 mg/L

TEST CONCENTRATIONS
Initial mean measured concentration: 0.077 mg/I
Mean measured concentration after 24h exposure: 0.031 mg/I
Mean measured concentration after 96h exposure: 0.019 mg/I

Reference substance (positive control):

yes

Remarks:

Pentachlorophenol

Dose descriptor:

NOEC

Effect conc.:

0.03 mg/L

Nominal / measured:

estimated

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Duration not specified

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 0.03 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 0.03 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No fish died during the total exposure period and no clinical effects were observed. In addition, no undissolved test material was noted.

Results with reference substance (positive control):

- Results with reference substance valid? Yes

- Mortality:
Under the conditions of the present test PENT ACHLOROPHENOL induced no lethal effects in carp at or below 0.15 mg/I.


- LC50:
The 96h-LC50 for carp exposed to PCP was 0.21 mg/I (95 % confidence interval between 0.19 and 0.27 mg/I).
The 24h-LC50 was 0.24 mg/I (95% confidence interval between 0.21 and 0.30 mg/I), and remained unchanged until 72h.
The range of the 96hLC50 for carp is generally between 0.10 and 0.46 mg/I based on historical data of reference tests performed approximately every 3 months from April 1988 until the end 2000, and annually since then. The response observed in carp originating from the present batch falls within this range.

Validity criteria fulfilled:

yes

Remarks:

The study met the acceptability criteria prescribed by the protocol.

Conclusions:

Setafix X 11342 induced no visible effects in carp exposed to a filtrate prepared at a loading rate of 100 mg/L. The average exposure concentration was 0.030 mg/L (NOEC), which was above water solubility.

The 96h-LC50 was above wateir solubility. Hence, due to the extremely low water solubility of the test substance, concentration levels toxic for fish could not be reached.

Executive summary:

The purpose of the study was to evaluate the test substance for its ability to generate acute toxic effects in Cyprinus carpio during an exposure period of 96 hours and, if possible, to determine the LC50 at all observation times. The study was based on the ISO International Standard 7346-1: Static method, 1996 and also designed to meet the test methods and validity criteria of the EEC directive 92/69, Part C.1, 1992 and the OECD guideline No. 203, 1992.

A limit test was performed exposing 7 fish 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 filteir (ca. 5 μm). The filtrate was clear and colourless. The total test period was 96 hours. Samples for analysis were taken at the start, after 24 hours and at the end of the test.

Analyses showed that the measured initial concentration was 0.077 mg/L. After 24 hours of exposure, the measured concentration was 0.031 mg/L. During the remainder of the exposure period the measured concentration decreased to 0.019 mg/I (26 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/L (NOTOX Project 330918).

Setafix X 11342 induced no visible effects in carp exposed to a filtrate prepared at a loading rate of 100 mg/L. The average exposure concentration was 0.030 mg/L (NOEC), which was above water solubility.

The 96h-LC50 was above wateir solubility. Hence, due to the extremely low water solubility of the test substance, concentration levels toxic for fish could not be reached.

Description of key information

1077 induced no visible effects in zebrafish exposed to a filtrate prepared at a loading rate of 100 mg/L.

This suggested that LC50 of the test substance were above this concentration level. Due to its soluble limit, the average exposure concentration of Z1077 was measured to be 0.994 mg/L. Due to the low water solubility, it was reasonable to infer that in natural conditions the test substance would not reach concentration levels toxic to fish.

Additional information

In one study the test substance was evaluated for its ability to generate acute toxic effects Z1077 in Brachydanio rerio during an exposure period of 96 hours and, if possible, to determine the LC₅₀ at all observation times. The study procedure was based on the OECD guidelines for Testing of Chemicals, guideline No. 203: "Fish Acute Toxicity Test", adopted 17 July, 1992. The test was performed by exposing 10 fish to a filtered solution prepared at a loading rate of 100 mg/L and a blank-control. The test solution was prepared by magnetically stirring of the dispersion for 8 hours, after which it was filtered over a paper filter. The total test period was 96 hours. Samples for analysis were taken at the start, after 24, 48, 72, and 96 hours after the exposure.

Chemical analysis showed that the measured initial concentration was 1.10 mg/L. After 24 hours of exposure, the measured concentration was 1.05 mg/L. At the end of the 96h exposure period the measured concentration decreased to 0.891 mg/L.

Z1077 induced no visible effects in zebrafish exposed to a filtrate prepared at a loading rate of 100 mg/L. This suggested that LC₅₀ of the test substance were above this concentration level. Due to its soluble limit, the average exposure concentration of Z1077 was measure to be 0.994 mg/L.

Due to the low water solubility, it was reasonable to infer that in natural conditions the test substance would not reach concentration levels toxic to fish.

In another study the test substance was evaluated for its ability to generate acute toxic effects in Cyprinus carpio during an exposure period of 96 hours and, if possible, to determine the LC50 at all observation times. The study was based on the ISO International Standard 7346-1: Static method, 1996 and also designed to meet the test methods and validity criteria of the EEC directive 92/69, Part C.1, 1992 and the OECD guideline No. 203, 1992.

A limit test was performed exposing 7 fish 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 96 hours. Samples for analysis were taken at the start, after 24 hours and at the end of the test.

Analyses showed that the measured initial concentration was 0.077 mg/L. After 24 hours of exposure, the measured concentration was 0.031 mg/L. During the remainder of the exposure period the measured concentration decreased to 0.019 mg/I (26 % of initial). Thus, the actual concentration was maintained above the water solubility, which was 8.8 μg/L (NOTOX Project 330918).

Setafix X 11342 induced no visible effects in carp exposed to a filtrate prepared at a loading rate of 100 mg/L. The average exposure concentration was 0.030 mg/L (NOEC), which was above water solubility.

The 96h-LC50 was above water solubility. Hence, due to the extremely low water solubility of the test substance, concentration levels toxic for fish could not be reached.