4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

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:

26 Oct - 09 Nov 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Health Care Inspectorate of the Ministery of Health, Welfare and Sport, Utrecht, The Netherlands

Test material

Test material information

Constituent 1

Reference substance name:

4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

EC Number:

235-985-8

EC Name:

4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

Cas Number:

13080-86-9

Molecular formula:

C27H26N2O2

IUPAC Name:

4,4'-[isopropylidenebis(4,1-phenyleneoxy)]dianiline

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: All test concentrations and the control
- Sampling method: Samples were taken at the start of the test and after 24 h from the freshly prepared solutions and at renewal (t= 24 h) and the end of the test from the 24 h old solutions. 2.0 mL were taken from the approximate centre of the test vessels.
- Sample storage conditions before analysis: not applicable

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Preparation of test solutions started with a loading rate of 100 mg/L applying a three-day period of magnetic stirring to ensure maximum dissolution of the test item in medium. Thereafter, the aqueous Saturated Solution (SS) was collected by filtration through a 0.45 µm membrane filter (RC55, Whatman) and used as the highest test concentration. Lower test concentrations were prepared by subsequent dilutions of the SS in test medium.
- Controls: Yes
- Eluate: No
- Differnetial loading: No
- Evidence of undissolved material: No; All test solutions were clear and colorless at the end of the preparation procedure.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: Straus, 1820
- Source: In-house laboratory culture with a known history
- Age of parental stock (mean and range, SD): 4 weeks
- Feeding during test: No feeding
- Food type: Suspension of fresh water algae
- Frequency: Daily

ACCLIMATION
- Acclimation period: 4 weeks
- Acclimation conditions (same as test or not): same
- Health during acclimation (any mortality observed): Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: < 24 hours, from parental daphnids of more than two weeks old

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg/L expressed as CaCO3

Test temperature:

20 - 21 °C

pH:

7.7 - 8.0 (Control)
7.7 - 8.0 (Test item concentrations)

Dissolved oxygen:

9.0 - 9.2 (Control)
8.5 - 9.1 (Test item concentrations)

Nominal and measured concentrations:

1.0, 10 and 100% of a SS prepared as a concentration of 100 mg/L
0.00040, 0.00078 and 0.0016 mg/L (measured concentration based on TWA)

Details on test conditions:

TEST SYSTEM
Test vessel:
- Type: open
- Material, size, headspace, fill volume: glass, 100 mL, headspace: 20 mL, fill volume: 80 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): After 7 days of cultivation half of the medium was replaced twice a week.
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4 (for highest test item concentration 100 mg/L) and 2 (for the other test item concentrations 1.0 and 10 mg/L)
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: M7 medium (adjusted ISO medium) according to the guideline
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH and dissolved oxygen were measured at the beginning, after 24 h and at the end of the test, for the control and the highest test concentration. Additionally, in the intermediate test concentrations upon renewal of the test solutions. The temperature was measured continuously in a temperature control vessel, beginning at the start of the test and in the fresh and spent test solutions upon renewal.

OTHER TEST CONDITIONS
- Photoperiod: 16 h photoperiod daily

EFFECT PARAMETERS MEASURED: Immobility (including mortality) was measured and recorded at 24 h and at 48 h.

RANGE-FINDING STUDY: no range-finding test was performed

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 0.002 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Remarks:

filtered solution

Basis for effect:

mobility

Details on results:

- Mortality of control: No
- Abnormal responses: No
- Others: Analytical and biological results are summarized in table 1 and 2 within the section "Any other information on results incl. tables"

Results with reference substance (positive control):

In conclusion, the actual responses in this reference test with K2Cr2O7 are within the ranges of the expected responses at the different concentrations, i.e. the 48 h EC50 was between 0.3 and 1.0 mg/L. Hence, the sensitivity of this batch of D. magna was in agreement with the historical data collected at Charles River Den Bosch. The 48 h EC50 was 0.41 mg/L with a 95% confidence interval between 0.34 and 0.46 mg/L.

Any other information on results incl. tables

Table 1: Analytical Results: Measured Concentrations

Test item 1 % SS prep. at 100 mg/L	Measured concentration (mg/L)				Average exposure concentration (mg/L)
	t= 0 h (fresh)	t= 24 h (old)	t= 24 h (fresh)	t= 48 h (old)
1.0	0.00040 2	0.00040 2	0.00040 2	0.00040 2	0.00040
10	0.00040 2	0.00040 2	0.00335	0.00040 2	0.00078
100	0.00354	0.00040 2	0.00332	0.00134	0.0016

1 4,4’-[isopropylidenebis(4,1-phenyleneoxy)]dianiline
2 Taken as half of the limit of detection, i.e. LOD/2 = 0.00040 mg/L.

Table 2: Biological Results: Number of Introduced Daphnids and Incidence of Immobility

Time (h)	Replicate	4,4’-[isopropylidenebis(4,1-phenyleneoxy)]dianiline % SS prep. at 100 mg/L
		Control	1.0	10	100
0	A	5	5	5	5
	B	5	5	5	5
	C	5			5
	D	5			5
	Total introduced	20	10	10	20
24	A	0	0	0	0
	B	0	0	0	0
	C	0			0
	D	0			0
	Total immobilised	0	0	0	0
	Effect %	0	0	0	0
48	A	0	0	0	0
	B	0	0	0	0
	C	0			0
	D	0			0
	Total immobilised	0	0	0	0
	Effect %	0	0	0	0

Table 3: Validity criteria for OECD 202

Criterion from the guideline	Outcome	Validity criterion fulfilled
In the control, including the control containing the solubilising agent, not more than 10% of the daphnids should have been immobilized.	No mortality	Yes
The dissolved oxygen concentration at the end of the test should be ≥ 3 mg/L in control and test vessels.	>3 mg/L	Yes

 

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Conclusions:

In conclusion, the 48 h EC50 for Daphnia magna exposed to 4,4’-[isopropylidenebis(4,1-phenyleneoxy)]dianiline was beyond the range tested, i.e. exceeded an average concentration of 0.0016 mg/L being considered the maximum soluble concentration of the test item in test medium at a concentration of 100 mg/L.