Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

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:

2017-09-05 - 2018-06-27

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: Ministry of environmental protection of the people's Republic of China, The Guidelines for the Testing of Chemicals-Effects on Biotic Systems, 2013, 211 Daphnia magna Reproduction Test

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

EC Number:

700-064-6

Cas Number:

2105830-60-0

Molecular formula:

Not applicable as this is a UVCB substance

IUPAC Name:

Esterification product of castor oil and tetrahydromethyl-1,3-isobenzofuranedione

Specific details on test material used for the study:

Testsubstance code: BLKN20170414-1787

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Water-soluble fractions (WSF) of the test substance were used as the test solutions. The WSFs were prepared by adding the required amount of the test substance into 5L test water. The mixtures were stirred in the dark for 72 hours. The vortex depth were 1/10 of the solution . The Water accommodated fraction (WSF) of 1 L was siphoned from the middle portion of the solution in the vessel after being settled for one hour (the first 100mL discarded), and then be filtered using a 0.45µm nitrocellulose membrane to prepare WSF as test solutions. The WSFs at each loading were prepared individually and 3 times a week.
The pH of the highest loading rate treatment group measured during the test was 8.161-8.331 , not exceeding 6-9, and needed not to be adjusted.
150ml samples of test water, spiked samples and test water blank were extracted with 75ml ethyl acetate by shaking at 220rpm for 10 minutes and standing for 10min. The ethyl acetate layer was concentrated to 1 ml at 30°C .Then it was transferred to 2ml sample vial with THF, and dried by nitrogen at 3o·c. Finally, the sample was diluted with THF to 1 ml and was analyzed directly by GPC.

Test solutions

Vehicle:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Daphnia magna is the species recommended in "Ministry of environmental protection of the people's Republic of China, The Guidelines for the Testing of
Chemicals-Effects on Biotic Systems, 2013, 211 Daphnia magna Reproduction Test" and "OECD, Guidelines for the Testing of Chemicals, 2012, 211 Daphnia magna Reproduction Test". Ease of culturing and handling, sensitivity to a variety of chemical substances, and the extensive data base for this species makes it suitable for acute toxicity testing .
Daphnia (ID: DM-BSAL-F2-171205-M4) were originally supplied by MicroBioTests Inc. Young female Daphnia (the parent animals) aged less than 24 hours and not first brood progeny were used at the start of the test. Adult Daphnia magna were bred in the laboratory of APM in the Elendt M4 medium. The culture conditions were similar to test conditions. Cultures of adult Daphnia magna were fed with concentrated suspensions of unicellular green algae Desmodesmus Subspicatus,
Pseudokirchneriella subcapitata and Ch/orella sp. In deinionized water at a rate equivalent to 0.1 mg-0.2mg of organic carbon per adult per day.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Test conditions

Hardness:

155 mg/L - 165 mg/L (as CaCO3)

Test temperature:

20.0 - 20.4 °C

pH:

8.015 - 8.753

Dissolved oxygen:

8.03 - 10.43 mg/L

Details on test conditions:

During the test period, the pH values of the highest test solution ranged from 8.015 to 8.951 , not exceeding 6-9 and not varying by more than 1.5 units; the dissolved oxygen concentrations of the test solutions ranged from 6.81 mg/L to 1 0.43mg/L, (recommended: above 3mg/L); the hardness of the test solutions ranged from 145mg/L to 165mg/L (as CaC03), (recommended: above 140mg/L (as CaC03)). During the test period , the Light intensity ranged from 12381ux to 13241ux, not exceeding 1000 Lux-1500 Lux; the temperature of the test media were between 20.0°C and 20.4°C, not exceeding 20°C±2°C and not varying by more than 2°C.
Each adult Daphnia magna was fed during the test at a rate equivalent to about 0.15mg of organic carbon per adult per day, not exceeding 0.1 mg-0.2mg of organic carbon.

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 10.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 2

Duration:

21 d

Dose descriptor:

LOELR

Effect conc.:

> 10.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Effect concentrations 3

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

>= 10.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Adult length

Effect concentrations 4

Duration:

21 d

Dose descriptor:

LOELR

Effect conc.:

> 10.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Adult length

Details on results:

The actual concentrations of the test substance in the test solutions were determined by GPC method. The limit of detection (LOD2) for the test substance was calculated to be 65.1ng/ml (based on a noise of 4.71) . Before sample analysis, the recovery of method was determined by blank and spiked samples. lt was found that the average recoveries were 1 0 2 % and 103% of the spiked 130ng/ml and 260ng/ml samples with RSD values of 0.76% and 2.06%, respectively. The concentrations of the test substance in the test solutions were analyzed by using GPC method. The measured concentration of the highest loading rate treatment group was below the limit of detection (LOD2) in the definitive test and the LOD2 was 65.1 ng/ml. The test results were expressed as the loading rates of the treatment groups. The mortality of the parent animals in the blank control group and in the treatment groups both were 0%, not exceed 20%. The mean number of live offspring produced by each parent animal surviving at the end of the test in blank control group was 111, not less than 60 and the coefficient of variation was 20.8%, not higher than 25%. According to the result of the method of Williams' Test, there was no significant difference in 21 days reproduction between the treatment group and the blank control (p=0.786 , p>0.05). The 21 days NOELR for reproduction of the test substance to Daphnia magna was 10.1 mg/L and the LOELR was higher than 10.1 mg I L. The adult length in each group at the end of the test are given in section 19.7. According to the result of the Dunnett's Test(p=0.929 , p>0.05), the 21 days NOELR for adult length of the test substance to Daphnia magna was 10.1 mg/L and the LOELR was higher than 10.1 mg/L.
No aborted egg and dead offspring were observed in each group during the test.

Results with reference substance (positive control):

Test(s) with a reference substance potassium dichromate were done regularly. The acute effect of the toxic reference substance potassium dichromate was tested at five concentrations of 0.320, 0.560, 1.00, 1.80, 3.20mg/L from 24 Aug. 2017 to 25 Aug. 2017 (24 hours test). The test organisms were immobilized by the potassium dichromate solutions by 0% to 85% (see Appendix 3). The percentages immobilised at 24 hours are plotted against test concentrations. Data are analysed by Maximum Likelihood-Probit to calculate the slopes of the curves and the EC50 with 95% confidence limits using software Toxcalc v5.0.32. The 24h
EC50 for potassium dichromate was 1.72mg/L, within accepted limits of ISO 6341 (the 24h EC50 for potassium dichromate should be between 0.6mg/L and 2.1 mg/L), and therefore the sensitivity of the daphnids meets the requirements.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Results showed that under the testing conditions, for Daphnia magna reproductions, the No Observed Effect Loading Rate (NOELR) was 10.1 mg/L, and the 50% reduction in reproductive output (EL50) greater than 10.1 mg / L. For Daphnia magna length, the No Observed Effect Loading Rate (NOELR) was 10.1 mg/L.
The concentrations of the test substance in the test solutions were analyzed by using GPC method. The measured concentrations of the highest concentration treatment group were below the limit of detection (LOD2) in the definitive test and the LOD2 was 65.1 ng/ml. The test results were expressed as loading rates of the treatment groups.

Executive summary:

Results showed that under the testing conditions, for Daphnia magna reproductions, the No Observed Effect Loading Rate (NOELR) was 10.1 mg/L, and the 50% reduction in reproductive output (EL50) greater than 10.1 mg / L. For Daphnia magna length, the No Observed Effect Loading Rate (NOELR) was 10.1 mg/L.

The concentrations of the test substance in the test solutions were analyzed by using GPC method. The measured concentrations of the highest concentration treatment group were below the limit of detection (LOD2) in the definitive test and the LOD2 was 65.1 ng/ml. The test results were expressed as loading rates of the treatment groups.