Reaction mass of (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl palmitate and (9-acetoxy-3,8,10-triethyl-7,8,10-trimethyl-1,5-dioxa-9-azaspiro[5.5]undec-3-yl)methyl stearate

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:

04 NOVEMBER 2015 - 06 NOVEMBER 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 110 mg/L WAF
Sampling at 0 h, 24 h and 48 h before and after the renewal of the test medium

The test solutions of 110 mg/L-WAF were filtered through a 0.22 µm filter (discarded the first 5 mL to 10 mL of the filtrations), and then the filtrations were filtered with methanol twice
before analysis. The blank controls were filtered through a 0.2 µm filter (discarded the first
5ml to 10 mL of the filtrations) before analysis. The samples were analyzed immediately after
sampling.

Vehicle:

no

Details on test solutions:

The test substance which is a complex liquid mixture and contains 60.9% of C-18, 30.7% of C-16, and 8.4% of other unknown constituents.
The solubility of test substance was quite poor, and the constituents C-18 and C-16 of the test substance can degrade quickly in water to the two degradation products CG40-0354 (CAS 376588-04-4) and CG43-0244 (CAS 60130-68-9) . Please see also the attached ilustration "Hydrolysis product of EC 936-831-9".

For the purposes of the test, the test solution was prepared as a Water Accommodated Fraction (WAF), and the method of preparing test solution was as follows:
0.33g test substance was filled in a 5L-jar and diluted with test water to 3000mL. The test suspension was stirred with a magnetic stirrer for about 20h (the depth of vortex was maintained in the range of 1cm-2cm), to obtain the nominal loading rate of 110 mg/l suspension. After letting the solution stand for about 1 hour when the stirring is over, the suspension in the middle layer was siphoned and the nominal loading rate of 110 mng/L WAF (Water Accommodated Fraction) was obtained. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test substance to be present, and it was directly used in the test.

In a pretest, the water solubility was evaluated in M4 media: The solubility test showed that test substance was poorly dissolved in test water. The
measured concentrations of the parent compound (CG 40-0946 and CG 43-0312) in the 114 mg/L loading rate suspension were lower than the LOD (LOD of CG40-0946 = 0.0826mg/L, LOD of CG43-0312 = 0.0377mg/L) after stirring 48h.

Test organisms (species):

Daphnia magna

Details on test organisms:

(1) Species: Daphnia mnagna
(2) Reason for selection of species: The test species is recomminended by "The Guidelines for the Testing of Chemicals".
(3) Source: Safety Evaluation Center, Shenyang Research Institute of' Chemical Industry. Apparently healthy, bred in our laboratory. The introduction batch number of test organism was DM20130801 -1, and the acclimated batch number of test animal was DMU20160120-l.
(4) Breeding and holding: At the start of the test, Daphniua magna were acclimated in the testing laboratory for 48h at
least under the same conditions of the test condition (water quality: Elendt M4 medium, temperature: 20°C±2°C, light and dark period: 16h light / 8h dark, illumination intensity:1000 Is -1500 Ix). During the acclimation, Daphnia maguna were fed with a concentrated algal suspension.
(5) Daphnia age: The animals were less than 24 hours old and were not first brood progeny.
(6) The number of test daphnia: 40 animals in total
(7) Sensitivity: The result of sensitivity test showed that 24h-EC, of reference substance K2Cr2O 7 is 1 .25mg/L within the range of 0.6 mg/L to 2.1 mg/L. And the 95% confidence limits is 1.13mg/L to 1.38mg/L.

Test type:

semi-static

Water media type:

freshwater

Remarks:

renewal every 24 hours

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

20.1 °C to 20.5 °C

pH:

see table 1 in "Any other information on materials and methods incl. tables"

Dissolved oxygen:

see table 1 in "Any other information on materials and methods incl. tables"

Nominal and measured concentrations:

WAF 110 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 150 mL glass beaker
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume:
- Volume of solution: 50 mL
- Aeration: no

- Renewal rate of test solution (frequency/flow rate): 24 h
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): -
- Biomass loading rate: 1 daphnid/10 mL solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h dark
- Light intensity: 1120 - 1123 lx

Range finding test:

Concnetrations: 12 mg/L, 32 mg/L and 116 mg/L WAF
duration: 48 h
Replicate: 2/test group
Daphnids: 5/repliacte, 40 in total
Testdesign: Static

Reference substance (positive control):

yes

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

>= 0.861 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

other: Hydrolysis product

Basis for effect:

mortality

Remarks on result:

other: Based on degradation product CG 40-0354

Details on results:

The measured concentrations of degradation products (CG40-0354 and CG43-0244) in the blank control at 0h, 24h and 48h before and after the renewal were both lower than the limit of detection (LOD) 0.000401 mg/L and 0.00107mg/L.
The measured concentrations of degradation product (CG40-0354) in the loading rate
110mg/L WAF test group at Oh, 48h and 24h before and after the renewal were 0.862mg/L (Oh new), 0.720mg/L (24h old), 0.878mg/L (24h new) and 1.01mg/L (48h old). The measured concentrations of degradation product (CG43-0244) in the loading rate 110mg/L WAF test group at Oh, 48h and 24h before and after the renewal were both lower than the limit of detection (LOD) 0.00107mg/L.

The test results revealed that immobilities of blank control at 24h and 48h after the start of the test were 0%, which were below than 10% and met the requirements of the validity criteria. Immobilities of the loading rate 110mg/L WAF test group at 24h and 48h after the start of the test were 0%.

Results with reference substance (positive control):

The result of sensitivity test showed that 24h-EC, of reference substance K2Cr2O 7 is 1 .25mg/L within the range of 0.6 mg/L to 2.1 mg/L. And the 95% confidence limits is 1.13mg/L to 1.38mg/L.

Validity criteria fulfilled:

yes

Conclusions:

(1) At the end of the test, immobilites of blank control were 0%, which were below than 10% and met the requirement.
(2) During the test, the dissolved oxygen concentration in the test solutions during the test ranged from 9.0mg/L to 9.4mg/L, it was kept above 3 mg/L.
(3) The 24h-EC50 of reference substance K2Cr207 was 1.25mg/L with the 95% confidence limits of 1.13mg/L-1.38mg/L. The 24h-EC50 of K2Cr207 was within the range of 0.6mg/L-2.1mg/L.

Executive summary:

No acut effects have been observedf within the range of water solubility.

 

Description of key information

With high probability acutely not harmful to aquatic organisms. No toxic effects were observed within the range of solubility.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

>= 110 mg/L

Additional information

Thereare two studies available regarding acute toxicity to aquatic invertebrates. The key study was done with the test item EC 936-831-9, the supporting study was done with the structural analogue just containing C-18 carbon chain subunits. Both tests resulted in EC50 values > 100 mg/L WAF.