2,2'-(1,4-phenylene)bis[4-[(4-methoxyphenyl)methylene]oxazol-5(4H)-one]

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2,2'-(1,4-phenylene)bis[4-[(4-methoxyphenyl)methylene]oxazol-5(4H)-one]

EC Number:

257-055-0

EC Name:

2,2'-(1,4-phenylene)bis[4-[(4-methoxyphenyl)methylene]oxazol-5(4H)-one]

Cas Number:

51202-86-9

Molecular formula:

C28H20N2O6

IUPAC Name:

2,2'-(1,4-phenylene)bis[4-(4-methoxybenzylidene)-1,3-oxazol-5(4H)-one]

Test material form:

solid: particulate/powder

Details on test material:

Expiry date
2026-04-19 (Statement of producer)

Specific details on test material used for the study:

Test item Polysynthren-Gelb GG

Chemical Name 2,2’-(1,4-Phenylene)bis[4-[(4-methoxyphenyl)methylene]oxazol-
(EINECS, REACH) 5(4H)-one]

Batch number DEB2 259062 ag

C.I.Index Solvent Yellow 133 (S.Y. 133)

CAS number 51202-86-9

EINECS / EC number 257-055-0

Purity (certified) 93.1% (w/w)
98.4% (w/w) including isomer

Molecular formula C28H20N2O6

Molecular weight 480.48 g/mol

Water solubility < 0.02 mg/L (20 °C)

Appearance Yellow powder

Stability Stable in water and light at room temperature

Expiry date 2026-04-19 (Statement of producer)

Recommended storage Room temperature, keep container tightly closed and dry in a cool, well-ventilated place




Test Facility Actions

Receipt 2016-05-12

Identification parameter Name, batch number, state, consistency and color

Retention sample At least 1 g has been retained on 2016-05-27 and stored at 6 ± 2 °C.

Storage conditions Room temperature, protected from light in the tightly closed original container

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

Limit loading level
A saturated solution with a nominal loading of 1.00 mg/L of the test item was tested.

Preparation of the saturated solution
The saturated solution (1.00 mg/L test item was weighed out) was
prepared with dilution water (see Table 2) two days before the start of the exposure (at -48 hours).
The test item was mixed with the dilution water by constant stirring with a magnetic stirrer at approximately 1100 rpm for 48 hours at room temperature. After completion of stirring, undissolved particles were removed by membrane filtration (membrane filter 0.45 µm, RC, MACHEREY-NAGEL). The filter was saturated in order to avoid adsorption during the filtration. The first 25 mL of the filtrate were discarded. The filtration was interrupted for 15 minutes to allow for adsorption and saturation of the filter material with dissolved test item. Thereafter, the filtration was continued. The next 25 mL were discarded. The following filtrate, i.e. the saturated solution, was used as a limit concentration in the test. During filtration, the filter was always kept covered.

Control
Dilution water without test item incubated under the same conditions as the test group

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system
Daphnia magna STRAUS (Clone 5)

Reason for the selectionof the test system
Daphnia magna is the preferred species in accordance with the
test guideline and is bred at the test facility.

Origin
Institut für Wasser-, Boden- und Lufthygiene (WaBoLu),
14195 Berlin, Germany

Breeder
Noack Laboratorien GmbH,
Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany

Culture
In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20  2 °C, in an incubator, 16 hours illumination; light intensity of max. 1500 lx

Culture medium
Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used. The composition of the culture medium is presented in Table 2.

Culture feeding
The culture daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL. The algae were cultured at the test facility.

Origin of the food algae
Sammlung von Algenkulturen (SAG),
Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Total hardness [mg CaCO3/L] : 180

Test temperature:

2016-09-13: 20.5 °C

pH:

Water Quality Parameters at the Start of the Exposure (0 hours)
(measured in one additional replicate of the saturated solution and the control)

Nominal loading level
of the test item
[mg/L] pH-value Dissolved O2 concentration [mg/L]
1.00* 7.30 7.01
Control 7.45 8.28
* = saturated solution


Water Quality Parameters at the End of the Exposure (48 hours)
(measured in all replicates of the saturated solution and the control)
Nominal loading level
of the test item
[mg/L] pH-values Dissolved O2 concentration [mg/L]]
Replicates Replicates
1 2 3 4 1 2 3 4
1.00* 7.25 7.30 7.33 7.33 7.32 7.12 7.75 7.96
Control 7.18 7.17 7.18 7.22 7.43 7.24 7.13 6.89
* = saturated solution

Dissolved oxygen:

Water Quality Parameters at the Start of the Exposure (0 hours)
(measured in one additional replicate of the saturated solution and the control)

Nominal loading level
of the test item
[mg/L] pH-value Dissolved O2 concentration [mg/L]
1.00* 7.30 7.01
Control 7.45 8.28
* = saturated solution


Water Quality Parameters at the End of the Exposure (48 hours)
(measured in all replicates of the saturated solution and the control)
Nominal loading level
of the test item
[mg/L] pH-values Dissolved O2 concentration [mg/L]]
Replicates Replicates
1 2 3 4 1 2 3 4
1.00* 7.25 7.30 7.33 7.33 7.32 7.12 7.75 7.96
Control 7.18 7.17 7.18 7.22 7.43 7.24 7.13 6.89
* = saturated solution

Conductivity:

2016-09-13 [µS/cm] : 500

Details on test conditions:

Test method
The study was performed with a static test design.

Test duration
48 hours

Test vessels
Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses

Test volume 2
0 mL in each test replicate (see ‘Application’)

Dilution water
Same as culture medium (see Table 2)

Number of daphnids and replicates
20 daphnids, divided into 4 replicates, each with 5 daphnids, were
used for the saturated solution and the control.

Age of the daphnids at the start of the exposure
Less than 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded.
The juveniles born within the following period of max. 24 hours preceding the exposure were used for the test. No first brood progeny was used for the test.

Acclimatization
Acclimatization of the daphnids was not necessary, because the dilution water was equivalent to the culture medium.
 
Application
20 g test solution per replicate was weighed out into each test vessel. This corresponds to 20 mL per test vessel. The daphnids were inserted with a small amount of dilution water by a pipette.

Test temperature (target)
18 - 22 °C, constant within ± 1 °C

Illumination (target)
Diffuse light, light intensity of max. 1500 lx

Photoperiod (target)
16/8 hours light/dark cycle

Feeding
The daphnids were not fed during the study.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

other: EL 10/50/100

Effect conc.:

> 1 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

other: EL 10/50/100

Effect conc.:

> 1 mg/L

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

The percentage of immobility for the reference item was determined after 24 hours. The EC50-value with 95% confidence limits (CI) was calculated by sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was:
EC50: 2.03 mg/L (Cl: 1.17 - 3.63 mg/L)
The EC50-value of the reference item potassium dichromate after 24 hours is within the required concentration range of 0.6 - 2.4 mg/L of quality criteria according to AQS P 9/2 (02/2000) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Any other information on results incl. tables

Methods of evaluation        

 The effects of the saturated solution were deduced directlyfrom the observed immobilization rates.
Alleffect levels (EL_{10 / 50 / 100}) given were based on the nominal test item loading.

 

EC-values and statistical  

analyses for reference item  

 

An EC₅₀-value was calculated for the reference item by sigmoidal dose-response regression.The respective 95% confidence limits were calculated from the standard error and the t-distribution. All calculations were carried out from the best-fit values with the software GraphPad Prism5.

 

Software                                 

All data were computer-processed and rounded for presentation. Consequently, minor variations may occur from the original figures if manual calculations based on the original figures are made subsequently. Calculations were made using the following software:

                                              - GraphPad Prism5,GraphPad Software, Inc.

                                              - Excel,Microsoft Corporation

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the saturated solution with a nominal loading of 1.00 mg/L of the test item Polysynthren-Gelb GG, no effects on Daphnia magna were observed.

Executive summary:

In an acute immobilization test with Daphnia magna (STRAUS), the effects of a saturated solution with a nominal loading of 1.00 mg/L of the test item were determined at the test facility according to OECD 202 (2004) from2016‑09‑11 to 2016-09-15, with the definitive exposure phase from 2016-09-13 to 2016-09-15.

The limit test was conducted under static conditions over a period of 48 hours.The saturated solution was colorless and visually clear during the exposure period. Twenty daphnids, divided into 4 replicates with 5 daphnids each, were exposed to the saturated solution and the control.

No analytical monitoring was performed during the test period in agreement with the sponsor, since no suitable method for the determination of the test item could be established. The test item is insoluble in water and organic solvents which are compatible for liquid chromatography.Exposures are expressed in terms of the nominal concentration of the test itemPolysynthren-GelbGG.

The validity criteria of the test guideline were fulfilled.

 

   EL_{10 / 50 / 100}-Values

(based on the nominal loading of the test item)

Effect values	Test duration [h]	Nominal test item loading [mg/L]
EL10 / 50 / 100	24	> 1.00
	48	> 1.00

 

 

 

In the saturated solution with a nominal loading of 1.00 mg/L of the test item no effects onDaphnia magnawere observed.