4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with trimethylhexane-1,6-diamine

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:

21 March 2018 - 09 June 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with trimethylhexane-1,6-diamine

EC Number:

500-332-0

EC Name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with trimethylhexane-1,6-diamine

Cas Number:

153195-44-9

Molecular formula:

(C15 H16 O2 . C9 H22 N2 . C3 H5 Cl O)x

IUPAC Name:

4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with trimethylhexane-1,6-diamine

Test material form:

liquid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: Control, 0.10, 0.32, 1.0, 3.2 and 10 mg/L loading rate WAF
- Sample storage conditions before analysis: The samples were stored frozen prior to analysis.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Nominal amounts of test item (5.0, 16 and 50 mg) were each separately added to the surface of 5 liters of test water to give the 1.0, 3.2 and 10 mg/L loading rates respectively. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixtures allowed to stand for 1-Hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. The aqueous phase or WAF was removed by mid depth siphoning (the first 75 to 100 mL discarded). Microscopic inspection of the WAFs showed no micro dispersions of test item to be present. Dilutions were made from the 1.0 mg/L loading rate WAF to give further test concentrations of 0.10 and 0.32 mg/L loading rate WAF.
The concentration and stability of the test item in the test preparations were verified by chemical analysis at 0 and 48 hours.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: clone 5, originally supplied by University of Sheffield/UK
- Source: in-house laboratory cultures
- Feeding during test: no

ACCLIMATION
- Acclimation conditions: Elendt M7 medium
- Type and amount of food: Each culture was fed daily with a mixture of algal suspension (Desmodesmus subspicatus) and Tetramin® flake food suspension.

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES:
Gravid adults were isolated the day before initiation of the test, such that the young daphnids produced overnight were less than 24 hours old. These young were removed from the cultures and used for testing. The diet and diluent water are considered not to contain any contaminant that would affect the integrity or outcome of the study.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Test temperature:

18 to 22 °C with a maximum deviation of ±1 °C

pH:

8.0-8.6 at test start, 8.1 at test end

Dissolved oxygen:

8.8-8.9 mg O2/L at test start; 8.7-9.0 mg O2/L at test end

Nominal and measured concentrations:

0.10, 0.32, 1.0, 3.2 and 10 mg/L loading rate WAF

Details on test conditions:

TEST SYSTEM
- Test vessel: 150 mL beakers containing approximately 100 mL of test preparation
- Aeration: no
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light / 8 h darkness with 20 minute dawn and dusk transition periods
- Light intensity: 675 to 740 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
immobilization or adverse reactions to exposure were recorded at 24 and 48 hours after the start of exposure

RANGE-FINDING STUDY
- Test concentrations: Control, 0.01, 0.10, 1.0, 10 and 100 mg/L loading rate WAF
- Results used to determine the conditions for the definitive study:
No sub-lethal effects of exposure were observed throughout the test.
Based on the results of the range-finding test the following loading rates were assigned to the initial experiments: 1.0, 3.2, 10, 32 and 100 mg/L. Four experiments were conducted.
The results of the first, second and fourth experiments showed significant immobilization throughout all test concentrations. In the third experiment, more than 10% of the control group daphnids were immobilized.
The results of the first three initial tests failed to identify the EL50 value, therefore the fourth test was conducted using a series of test concentrations of 0.10, 0.32, 1.0, 3.2 and 10 mg/L.

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Results and discussion

Effect concentrations

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

0.64 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

loading rate WAF

Basis for effect:

mortality

Remarks on result:

other: 95% c.l.

Remarks:

0.43-0.94 mg/L loading rate WAF

Details on results:

- Other biological observations: Reduced mobility was observed in the test concentrations 0.10, 0.32, 1.0 and 3.2 mg/L loading rate WAF; pale appearance was observed in the 10 mg/L loading rate WAF test concentration
- Mortality of control: no
- Other adverse effects control: no
- Effect concentrations exceeding solubility of substance in test medium:
At the start of mixing the 1.0 mg/L loading rate WAF was observed to have formed a clear colorless water column with test item floating at the surface. The 3.2 and 10 mg/L loading rate WAFs were observed to have formed clear colorless water columns with test item floating at the surface and dispersed throughout. After stirring, and following a 1-Hour standing period, the 1.0 mg/L loading rate WAF was observed to have formed a clear colorless water column with test item floating at the surface. The 3.2 and 10 mg/L loading rate WAFs were observed to have formed clear colorless water columns with test item floating at the surface and on the bottom of the water column. Microscopic examination of the WAFs showed there to be no micro-dispersions of test item present.
During the test the control and all test solutions were observed to be clear, colorless solutions.

Results with reference substance (positive control):

- Results with reference substance valid?
yes
24 h EC50 = 0.79 mg/L, 95% c.l. 0.73-0.86 mg/L
48 h EC50 = 0.75 mg/L, 95% c.i. 0.56-1.0 mg/L

The results from the positive control with potassium dichromate were within the normal range for this reference item. The results of the inter laboratory tests and a Technical Corrigendum to ISO 6341 give a 24-Hour EC50 for potassium dichromate within the range 0.6 mg/L to 2.1 mg/L.

Reported statistics and error estimates:

The EL50 values and associated confidence limits at 24 and 48 hours and the slope of the response curve and its standard error were calculated by Probit analysis using Linear Maximum Likelihood regression. The Lowest Observed Effect Loading (LOEL) rate and the NOEL rate at 24 and 48 hours were calculated using the Step-down Cochran-Armitage Test Procedure. All results were calculated using the ToxRat Professional computer software package (TOXRAT).

Any other information on results incl. tables

Validity criteria

The test was considered to be valid given that none of the control daphnids showed immobilization or other signs of disease or stress and that the oxygen concentration at the end of the test was equal to or greater than 3 mg/L in the control and test vessels.

Cumulative Immobilization Data and Observations in the Definitive Test

Nominal Loading Rate (mg/L)	24 Hours
	Cumulative Immobilized Daphnia (Initial Population: 5 Per Replicate)						Observations
	Rep1	Rep2	Rep3	Rep4	Total	%	Rep1	Rep2	Rep3	Rep4
Control	0	0	0	0	0	0	5N	5 N	5 N	5 N
0.10	0	0	0	0	0	0	5 N	5 N	5 N	5 N
0.32	0	0	0	0	0	0	5 N	5 N	5 N	5 N
1.0	2	2	1	0	5	25	3 N	3 N	1 R 3 N	5 N
3.2	3	5	4	0	12	60	2 R	AI	1 R	5 N
10	2	3	5	5	15	75	3 P	2 N	AI	AI
Nominal Loading Rate (mg/L)	48 Hours
	Cumulative Immobilized Daphnia (Initial Population: 5 Per Replicate)						Observations
	Rep1	Rep2	Rep3	Rep4	Total	%	Rep1	Rep2	Rep3	Rep4
Control	0	0	0	0	0	0	5 N	5 N	5 N	5 N
0.10	0	1	1	0	2	10	5 N	4 N	1 R 3 N	5 N
0.32	1	1	0	2	4	20	4 N	4 N	2 R 3 N	3 N
1.0	4	3	3	1	11	55	1 R	2 R	1 R 1 N	3 R 1 N
3.2	5	5	5	5	20	100	AI	AI	AI	AI
10	5	5	5	5	20	100	AI	AI	AI	AI

 

P       = Pale

Rep     = Replicate

R         = Reduced mobility

N        = Normal

AI        = All daphnia immobilized

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Exposure of the freshwater invertebrate Daphnia magna to the test item has been investigated and gave the following results: 48 h EL50 = 0.64 mg/L loading rate WAF (95% Confidence Limits 0.43 - 0.94mg/L loading rate WAF).

Executive summary:

A study was performed to assess the acute toxicity of BADGE-TMD adduct to Daphnia magna. The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (April 2004) No 202, "Daphniasp., Acute Immobilisation Test" referenced as Method C.2 of Commission Regulation (EC) No. 440/2008.

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

Following a preliminary range‑finding test and initial experiments, 20 daphnids (4 replicates of 5 animals) were exposed to WAFs of the test item over a range of nominal loading rates of0.10,0.32,1.0,3.2and10 mg/L for 48 hours at a temperature of 21 °C to 22 °C under static test conditions. The number of immobilized daphnia and any adverse reactions to exposure were recorded after 24 and 48 hours.

Chemical analysis of the fresh test preparations at 0 hours showed measured test concentrations to range from 0.27 to 12 mg/L. Chemical analysis of the aged test preparations at 48 hours showed measured test concentrations to range from 0.28 to 12 mg/L.

The dissolved test item may have been one or several components of the test item. Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

Exposure of the freshwater invertebrate Daphnia magna to BADGE-TMD adduct has been investigated and gave the following results: 48 h EL50 = 0.64 mg/L loading rate WAF (95% Confidence Limits 0.43 - 0.94mg/L loading rate WAF).