Reaction mass of bis[4-(vinyloxy)butyl] hexane-1,6-diylbiscarbamate and 9,18-dioxo-3,8,19-trioxa-10,17-diazapentacos-1-en-25-yl [6-({[4-(vinyloxy)butoxy]carbonyl}amino)hexyl]carbamate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

03 Dec 2015 - 18 Jan 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2012

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: SEPA 2004, HJ/T 153, The guidelines for the Testing of Chemicals

Version / remarks:

2004

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: CRC-MEPT, The guidelines for the Testing of Chemicals, Effects on Biotic Systems

Version / remarks:

2013

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: GB/T 21828-2008, Chemicals-Daphnia magna reproduction test

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.20 (Daphnia magna Reproduction Test)

Version / remarks:

2008

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1300 (Daphnid Chronic Toxicity Test)

Version / remarks:

1996; EPA 712-C-96-120

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations: blank control; 100% of stock solution
- Sampling method: 10.0 mL water samples were taken from all control and test solutions at the beginning (0 d), immediately prior to the first renewal of the test solution (3 d), and weekly thereafter (9, 21 d).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Stock solution preparation: a supersaturated solution was prepared by adding 10.000 g test substance into 1 L Elendt M7 and shaking for 24h in the incubator (temperature: 35°C; rotor speed: 100 (±10) r/min rotor speed) and then leaving it to settle for 1h at room temperature. Then the aqueous portion above was filtered by GF/C membrande. The filtrated solution will be regarded as the stock solution.
- Test solution preparation: test solutions at different concentrations (in the range-finding test) were prepared by adding the stock solutions and diluting with Elendt M7.
- Controls: Blank control (Elendt M7 without test substance)

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain/clone: not specified
- Source: obtained from Nanjing University in 2010
- Feeding during test: yes
- Food type: Chlorella vulgaris Beij.
- Amount: 0.1-0.2 mg C/Daphnia
- Frequency: once every day
- Stock animals were maintained in culture conditions similar to those to be used in the test, Lot No: D20151202D.
- Daphnia used in the test were not first brood progeny, and were 6-24h old at the start of the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

221 - 232 mg CaCO3/L

Test temperature:

20.2 - 21.0 °C

pH:

7.46 - 7.80

Dissolved oxygen:

90 - 101% of air saturation at test temperature.

Nominal and measured concentrations:

Nominal: 100% stock solution
Measured concentration: 285 ug/L (mean, N=6). See Table 1 in 'Any other information on results' for details.

Details on test conditions:

TEST SYSTEM
- Test vessel: 100 mL glass beakers with lid with holes.
- Aeration: no
- Renewal of test media: every 72 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M7, according to OECD211
- Culture medium different from test medium: no
- Intervals of water quality measurement: Oxygen concentration, temperature, hardness and pH values were measured once a week.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16:8 h light:dark cycle daily
- Light intensity: 1000 - 1500 lux

EFFECT PARAMETERS MEASURED:
- The number of living offspring was counted for each vessel daily. The offspring produced by each parent animal were preferably removed and counted daily from the appearance of the first brood to prevent them consuming food intended for the adult.
- Other parameters: time to production of first brood; number of broods per animal.

TEST CONCENTRATIONS
- Range finding study test concentrations: Control, 10% of stock solution, 100% of stock solution.
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

285 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

other: Reproduction and mortality

Details on results:

- Analytical results showed that the concentration of the test substance was consistent in the test medium throughout the test period (deviation within 20% of the initial concentration). Thus a semi-static method of 72 h renewal test procedure was reasonable.
- Behavioural abnormalities: none observed
- Blank control and 100% stock solution treatment (285 ug/L): no mortality
- Mean fecundity (No. live offspring per adult): blank control: 76; 100% stock solution: 70; inhibition rate: 8.31%. There was no statistically significant difference between blank control and 100% stock solution reproduction (p > 0.05).

Reported statistics and error estimates:

Independent Samples T-test, SPSS 17.0

 Table 1: Measured Test Results of Test Substance in Test Media

Nominal concentration	Mean Measured Concentration (µg/L)*	Measured Concentration (µg/L)**
		Day 0	Day 3		Day 9		Day 21
		Fresh	Aged	Fresh	Aged	Fresh	Aged
Blank Control	ND***	ND	ND	ND	ND	ND	ND
100% stock solution	285	292	280	289	281	288	279

* The average of the analysed concentrations

** The concentration value in this table has considered 8 times dilution.

*** ND: not detected, since below the sensitivity of the analytical method

Validity criteria fulfilled:

yes

Remarks:

See 'Overall remarks' for details on validity criteria

Conclusions:

The NOEC for reproduction and survival of parent animals was 285 ug/L (mean measured concentration).

Executive summary:

The effect of the test substance on the reproductive output of Daphnia magna was assessed according to OECD Guideline 211 and Chinese national guidelines under semi-static conditions. In a limit test, 10 individually held Daphnia were exposed to a blank control and to a 100% stock solution treatment (filtrate of 1000 mg/L after 24h shaking and filtering). Mean measured concentration in the 100% stock solution treatment was 285 ug/L. No mortality was observed in blank control and 100% stock solution. The mean fecundity (No. live offspring per adult) was 76 for the blank control, and 70 for 100% stock solution (inhibition rate: 8.31%). There was no statistically significant difference between blank control and 100% stock solution reproduction (p > 0.05). The NOEC for both reproduction and survival was 285 ug/L (mean measured concentration). The test met all validity criteria and is considered reliable without restriction.

Description of key information

The effect of the test substance on the reproductive output ofDaphnia magnawas assessed according to OECD Guideline 211 and Chinese national guidelines under semi-static conditions. In a limit test, 10 individually heldDaphniawere exposed to a blank control and to a 100% stock solution treatment (filtrate of 1000 mg/L after 24h shaking and filtering). Mean measured concentration in the 100% stock solution treatment was 285 ug/L. No mortality was observed in blank control and 100% stock solution. The mean fecundity (No. live offspring per adult) was 76 for the blank control, and 70 for 100% stock solution (inhibition rate: 8.31%). There was no statistically significant difference between blank control and 100% stock solution reproduction (p > 0.05). The NOEC for both reproduction and survival was 285 ug/L (mean measured concentration). The test met all validity criteria and is considered reliable without restriction.

Additional information

NOEC: 285 ug/L, which is considered the maximum solubility of the test substance in test medium.