2-Butenedioic acid (Z)-, ester with 1,2-propanediol, compd. with 2-(dibutylamino)ethanol

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: According to international guideline(s), GLP-compliant, performed in recognized contract research organization, no restrictions, fully adequate for assessment.

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1010 (Aquatic Invertebrate Acute Toxicity Test, Freshwater Daphnids)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Chemical Substance Control Law 1973, amended 2009 under the reference of YAKUSHOKHATSU No. 1121002, SEIKYOKU No.2 and KANPOKIHATSU No. 021121002 and partially amended 2006 as the joint ordinance of Japanese METI, MHLW and MOE

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: nominal 100 , 50, 25, 12.5, 6.25 mg/L and control
- Sampling intervals: start and end of the renewal periods
- Sampling method: samples were taken from flasks containing no animals
- Sample storage conditions before analysis: no storage prior to analysis

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
The highest test concentration of 100 mg/L and dilutions thereof were prepared in test medium.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: National Institute of Public Health, 1097 Budapest Gyáli u. 2-6. Hungary. The Daphnia are bred in-house.
- Age at study initiation: < 24 h
- Feeding during test: none

ACCLIMATION
None; the same medium (ISO medium according to the OECD and EC guidelines) was used for breeding and tests. The health of the stock animals is continuously monitored by visual daily checking. Abnormal behavior or significant decrease of population is recorded.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

249 mg/L as CaCO3 (calculated)

Test temperature:

20.8-21.3 °C

pH:

7.48-7.54

Dissolved oxygen:

7.4-7.6 mg O2/L

Nominal and measured concentrations:

nominal concentrations: 100, 50, 25, 12.5, and 6.25 mg/L
freshly prepared test media:103-126% of nominal
aged test media (24 h): 111-137% of nominal (101-128% of initially measured)
The analytically measured concentrations deviated more than 20% from the nominal values in some cases, thus the biological results are based on the geometric mean measured concentrations: 114.3, 56.4, 28.0, 15.1, and 8.0 mg/L (112-128% of nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: open glass beakers containing approx. 25 mL medium
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: at least 5 mL medium per animal

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: deionised water
- Ca/Mg ratio: 4:1
- Culture medium different from test medium: no
- Intervals of water quality measurement: water temperature, oxygen concentration and pH in the controls and the test solutions were measured at the beginning and at the end of each renewal period.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16-hour light and 8-hour dark cycle

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : number of immobilised animals and percentage of immobility at 24 and 48 hours

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.0
- Range finding study
- Test concentrations: nominal100, 10, 1, and 0.1 mg/L
- Results used to determine the conditions for the definitive study: 20 and 10% immobility after 48 hours at 100 and 10 mg/L,respectively; no immobility at the lower test concentrations

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 114 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

28 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

56 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

- Number and percentage of immobiliosed daphnids: see Table 1
- Behavioural abnormalities: none
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none

Results with reference substance (positive control):

- Results with reference substance valid? yes
- 24-hour EC50: 0.98 mg/L (95% confidence limits. 0.92-1.06 mg/L)

Reported statistics and error estimates:

The 24-hour EC50 could not be calculated because of the lack of toxic effects. The 48-hour EC50 value was determined by Probit analysis using TOXSTAT software.
The 24-hour EC50, the 48-hour EC100, the LOEC and NOEC values were determined directly from the raw data.

Table 1: Number and percentage of immobilised daphnids

 

Concentration (mg/L) *	Cumulative number of immobile Daphnia
	24 hours						48 hours
	R1	R2	R3	R4	total	%	R1	R2	R3	R4	total	%
Control	0	0	0	0	0	0	0	0	0	0	0	0
8.0    (6.25)	0	0	0	0	0	0	0	0	0	0	0	0
15.1  (12.5)	0	0	0	0	0	0	0	0	0	0	0	0
28.0  (25)	0	0	0	0	0	0	0	0	0	0	0	0
56.4  (50)	0	0	1	0	1	5	1	1	1	1	4	20
114.3 (100)	0	0	0	1	1	5	1	1	1	1	4	20

*: geometric mean measured (nominal)

R: replicate number, 5 daphnids per replicate

 

Validity criteria fulfilled:

yes

Remarks:

In the control, no Daphnia have been immobilised or trapped at the surface of the water. The dissolved oxygen concentration at the end of the test was > 3 mg/L in control and test vessels.

Conclusions:

The results indicate that the test material is not toxic to daphnia (48-h EC50 > 100 mg/L).

Description of key information

48-h EC50 >100 mg/L for Daphnia magna (OECD 202, EU C.2)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

The results indicate that the test material is not toxic to daphnia up to the highest concentration required by the test guidelines and thus a PNEC cannot be determined.