Fatty acids, C8-10, diesters with 1,4:3,6-dianhydro-D-glucitol

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 31/05/10 to 09/11/10

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

yes

Principles of method if other than guideline:

The study was performed according to the approved Study Plan with the following changes:

1- The range-finding test was performed for test item treatment between approximately 0.03 and 20 mg/L instead of 1 and 1/10000 times the limit of solubility of the test item in water.
2- The full definitive test was performed with seven test item treatmens instead of five.
3- Full definitive test: the analytical check of the test concentration did not involve the lowest test item treatment because this was far below the quantification level for the analytical method. The analytical check was performed with the two highest test item treatments.
4- Food consisted in living cells of Chlorella sp. instead of Scenedesmus subspicatus.

In the opinion of the Study Director the above changes did not alter the quality and integrity of the study.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Isosorbide Diesters

EC Number:

700-073-5

Cas Number:

1215036-04-6

Molecular formula:

As it is an UVCB, we can not provide the exact molecular formula.

IUPAC Name:

Isosorbide Diesters

Details on test material:

- Name of test material (as cited in study report): Isosorbide Diesters

- Physical state:colourless translucent liquid
- Analytical purity: 97%
- Impurities : Mono-ester dianhydrohexitol 1%

- Lot/batch No.: CB 10 008

- Storage condition of test material: at room temperature protected from direct sun light

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

Young daphnids were exposed to the test substance at a range concentrations for a 21/22 days period. At the end of the test, the total number of living offspring produced per parent animal alive at the end of the test was assessed. The reproduction rate of specimens that were found dead during the exposure period was excluded from the analysis.
The reproductive output of the animals exposed to the test item was compared to that controls.
A first experiment was performed as a range-finding test using three replicates units for each of five test item concentrations between 0.04 and 24.25mg/L.
A second experiment was conducted as a full definitive test using ten replicate units for each of seven test item concentrations between 0.008 and 1.94mg/L.
The analytical verification of the treatment concentrations used for the full definitive test failed. It was suspected that adsorption of the test item onto SPE column was biased because of the presence of metasilicates in the mineral medium. Other experiments were attempted in modified mineral waters but all of them failed: the viability of the specimens was reduced and/or the reproduction rate of the control specimens was not sufficient with regards to the validity criteria.
Further investigation of the analytical method showed that temperature conditions above 20°C were responsible for that adsorption of the test item onto the SPE column was not total. It was thus concluded that the M4 mineral medium was convenient for testing and care was taken to maintain temperature conditions below 20°C for the analytical verification of the test item treatments.
Finally the test was conducted as a limit test at 1.0 mg/L to confirm that the test item had no induced effect at this threshold value, and to provide data concerning the stability of the test item concentrations throughout the exposure period.
Potassium dichromate was used as the reference item to confirm the function of the test system: the reproductive performance of the daphnids should be significantly reduced at 0.1mg/L.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

The strain was received from a commercial breeder and regularly sub-cultured at the Phytosafe site in M4 mineral medium. This M4 mineral was freshly reconstituted from pure water and aliquot samples of mineral stock solutions. Adult females were transferred in a new volume of water on the day before test initiation. Young daphnids produced within 24 hours were used for the tests.

Test Units : 60mL capacity EPA glass tubes filled with 50mL of reconstituted water.
One Daphnid per unit. Range-finding test : 3 replicate units for each test item treatment and for controls. Full definitive test : 10 replicate units for each test item treatment and for controls. Limit test : 15 replicates units for each test item treatment and for controls and 10 replicates units for each 0.1, 0.2 and 0.5mg Potassium dichromate per litre.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Test conditions

Details on test conditions:

In every case the test was performed 20.0 +/- 2.0°C and at 16 hours light/8 hours darkness and the test solutions were renewed at 2-3 day intervals, accounting for the week-end.
The parent animals were fed with living algal cells of chlorella at ration levels corresponding to 0.1-0.2 mg Carbon/Daphnia/day. A concentrated algal suspension was used to minimize the volume of algal culture medium transferred to the test medium. The food wad added after each renewal of the test solutions.
The offspring produced by each parent was removed and counted at 1-2 day intervals from the appearance of the first brood. The presence of dead offspring was recorded, if any. Other parameters that were measured or calculated included time to production of first brood, number of aborted broods, and presence of males or ephippia.
The number of dead adult animals was recorded.
Oxygen concentration, temperature, and pH values were measured at least once a week, in fresh and old media, in the control and in the highest test substance concentration, at least. The initial pH should be maintained in the range 6-9 and should not vary by more than 1.5 units in any single test.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate CAS 7778-50-9

Results and discussion

Effect concentrations

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Basis for effect:

mortality

Details on results:

The total offspring produced by parent animals alive at the end of test was calculated. The reproductive performance of the daphnids found dead during the exposure period was excluded from the analysis. Mean value and standard deviations were calculated for the controls and for each test group. F-variance analysis at the 5% confidence level was used to detect any significant difference between test and control groups.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The tests were considered as valid as the results fulfilled to the following conditions:
- In controls, the mortality of parents animals did not exceed 20% at the end of the test.
- The mean number of live offspring produced in controls per parent animal surviving at the end of the test was ≥ 60.
- The relative standard deviation fot the mean number of living offspring produced per parent animal in controls was ≤25%.
- The dissolved oxygen concentration at the end of the test was ≥3mg/L in control and test vessels/
- Data were provided to ascertain that the test concentration was satisfactorily maintained within 80-120% of the nominal value throughout the limit test for the 1.0mg/L test item treatment. For the full definitive test, the analytical assessments failed because of an unexpected technical bias in the analytical method. Thus valid verifications for the test item treatments were lacking.

It was thus concluded that the results of the full definitive test should not be considered in the final conclusions referring to the side-effect of the item on the reproductive performance of the daphnids.

Isosorbide Diesters had no induced effect on the reproduction of daphnids at 1.0 mg/L.
NOEC for reproduction ≥1.0 mg/L for isosorbide Diesters

Executive summary:

Daphnia magna reproduction test with Isosorbide diesters.

Test item : Isosorbide Diesters - Batch CB 10 008

Guidelines : OECD 211, October 2008

Nominal test concentrations :

Range-finding test: 0.04 - 0.19 - 0.97 - 4.85 and 24.25 mg/L              

Full definitive test: 0.008 - 0.02 - 0.05 - 0.12 - 0.31 - 0.78 and 1.94 mg/L

Limit test: 1.0 mg/L

Test dates: May 31 to November 9,2010

Length of test: 21 day

Test microorganisms: Daphnia magna

Source: PHYTOSAFE S.A.R.L. 2, rue Marx Dormoy. 64000 PAU. France

Age: Less than 24 hours at the test initiation

Control: water

Results:

Results	Water control	Solvent control	Isosorbide Diesters 1.0 mg/L
Mortality	13.3 %	6.7 %	13.3 %
Offspring production	82.8 +/- 16.2 (19.6%)	83.5 +/- 9.9 (11.8%)	81.5 +/- 14.7 (18.0%)
Conclusions	NOEC for reproduction ≥ 1.0 mg/L