Reaction products of 3-aminomethyl-3,5,5-trimethylcyclohexylamine and m-phenylenebis(methylamine) with 2,2'-[(1-methylethylidene)bis(4,1-phenyleneoxymethylene)]bisoxirane

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2008-04-15 to 2008-04-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study is regarded as reliable without restrictions because it was conducted in accordance with GLP regulation and guideline.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling method:
For determination of the test item concentration, samples were taken from the testing concentrations and the control at the start and at the end of the test.
- Sample storage conditions before analysis:
The samples were analysed directly after sampling. The samples will be kept stored in deep-freezer up to the date of the final report.

Vehicle:

no

Details on test solutions:

A stock solution with a concentration of 76.5 mg/L (measured; see Appendix 2, 2.3) was prepared with test item and ISO Medium by means of ultrasonic treatment two days before the start of the study (on day -2). The test solutions were prepared from this stock solution by the appropriate dilution with ISO Medium on day 0 as given in the table below. The stock solution and all test solutions were prepared without the use of organic solvents.
(In the main study four replicates were used for each test concentration and for the untreated control.)
Start of the test: immediately after preparation of the test solutions Daphnids were introduced (< 15 minutes).

Concentration (mg/L) Stock solution (mL) ISO Medium (mL) Total volume (mL)
nominal measured
Control - 300 300.00
0.171 0.136 0.67 300 300.67
0.376 0.288 1.48 300 301.48
0.826 0.705 3.27 300 303.27
1.818 1.569 7.30 300 307.30
4.0 4.6 16.55 300 316.55
8.8 9.4 39.00 300 339.00

Control:
In the control, test water was used without addition of the test item.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Straus) not first brood progeny
Age at Test Start: Less than 24 hours old
Sex: Female
Origin: National Institute of Public Health, Hungary
Breeding Conditions: The Daphnia were bred in the laboratories of LAB Research Ltd. under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test. The test animals were fed daily with a few mL of centrifuged algae suspension (Pseudokirchneriella subcapitata).
Acclimatisation: Brood Daphnids were maintained in ISO medium under test conditions prior to the start of the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

171.1 mg/L

Test temperature:

18 - 22 °C

pH:

8.2 - 8.35

Dissolved oxygen:

6.3 mg/L of higher

Nominal and measured concentrations:

nominal: 0.171; 0.376; 0.826; 1.818; 4.0 and 8.8 mg/L
measured: 0.136; 0.288; 0.705; 1.569; 4.6 and 9.4 mg/L

Details on test conditions:

Test Units
Type and Size: Glass beakers of 40 mL test medium
Identification: Each test unit was uniquely identified with study code, treatment and replicate number.

Test Conditions
Test Environment: Climate chamber
Temperature: 18 – 22 °C with a maximum deviation of ± 1 °C
Light period: 16 h light : 8 h dark, light intensity: 885 lux
Recording: Test conditions were measured with suitable instruments and documented in the raw data.

Test water
Reconstituted Water:
ISO medium (during the holding period and during the test). The hardness (CaCO3) of water was: 171.1 mg/L 1). 1) Determined by National Public Health and Medical Officer Service. Date of sampling: 15 April 2008; Results: 17 April 2008, The documentation is archived in the raw data notebook. The ISO medium (according to OECD 202) was prepared by adding 25 ml each of the following stock solutions to deionised water.
Substance
CaCl2 x 2 H2O 11.76 g/L
Batch No.: KBR5022501
Supplier: REANAL
Expiry date: May 2009
MgSO4 x 7H2O 4.93 g/L
Batch No.: A895286
Supplier: MERCK
Expiry date: 31 Aug. 2012
NaHCO3 2.59 g/L
Batch No.: 829880
Supplier: REANAL
Expiry date: Dec. 2007
NaHCO3 2.59 g/L
Batch No.: K37882029
Supplier: MERCK
Expiry date: 30 September 2012
KCl 0.23 g/L
Batch No.: PP/2007/09359/0
Supplier: REANAL
Expiry date: 31 May 2010

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.75 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

6.11 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.59 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

6.57 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

9.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

0.288 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.705 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

1.569 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

8.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

0.376 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.826 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

1.818 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Analytical Results
The analytically determined mean test item concentrations in the analysed test media varied in the range from 90 % to 115 % at the start and from 63 % to 117 % at the end of the study in comparison to the nominal value

Biological Results and Conclusion
Results after 48 hours:
The 48h-NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours) of Reaction products of IPDA with bisphenol A diglycidylether (BADGE) to Daphnia magna was determined to be 0.705 (nominal: 0.826) mg test item/L. The observed 5 % mortality at this concentration level is in the control data range. The 48h-LOEC was 1.569 (nominal 1.818) mg test item/L. While the 48h-EC50 was calculated to be 1.59 mg/L (nominal: 1.75 mg/L); the 48h-EC0 of the test item was 0.288 mg/L (nominal 0.376 mg/L) and the 48h-EC100 was determined to be 9.4 (nominal 8.8) mg test item/L.

Results with reference substance (positive control):

For the evaluation of the quality of the Daphnia clone and the experimental conditions, potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions. The date of the last study (Study Code: 07/541-023DA) with reference item was: 20-21 November 2007. The results of the reference study are the followings:

The 24h-EC50 value: 1.86 mg/L
95 % confidence limits: 1.50 – 2.21 mg/L

In the test with the reference item the results met the expected toxicity of the POTASSIUM DICHROMATE (the 24h EC50 value of the potassium dichromate should be within the range 0.6 mg/L to 2.1 mg/L).The results are based on the nominal concentrations.

Reported statistics and error estimates:

The EC50 values of the test item and their confidence limits were calculated using Probit analysis. The analysis was done using the statistical software program SPSS+. The EC0, the NOEC, the LOEC and the EC100 were determined directly from the raw data.

Validity criteria fulfilled:

yes

Conclusions:

In the 48-hour static acute toxicity test to Daphnia magna with Reaction products of IPDA with bisphenol A diglycidylether (BADGE) the EC50 was determined via Probit analysis to be 1.59 mg/L (nominal: 1.75 mg/L). The overall NOEC was determined to be 0.705 mg/L (nominal: 0.826 mg/L).

Executive summary:

The purpose of this study was to evaluate the influence of the test item Reaction products of IPDA with bisphenol A diglycidylether (BADGE) on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed in a static test to the test item for 48 hours, added to test water at a range of concentrations.

0.171; 0.376; 0.826; 1.818; 4.0 and 8.8 mg/L nominal test item concentrations and a control were tested in the main study. The corresponding mean measured test item concentrations were 0.136; 0.288; 0.705; 1.569; 4.6 and 9.4 mg test item/L.

The test item concentration of the test solutions was measured by HPLC-UV method at the start and at the end of the test.

The analytically determined test item concentrations in the analysed test media varied in the range from 90 % to 115 % at the start and from 63 % to 117 % at the end of the study in comparison to the nominal value. The deviation of the measured concentrations from the nominal values was higher than 20 % at the lowest two concentration levels at the end of the test. Therefore, all reported biological results are related to the geometric mean measured concentrations of the test item at the start and at the end of the test calculated by EXCEL Software Program.

The 48h-NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours) of Reaction products of IPDA with bisphenol A diglycidylether (BADGE) to Daphnia magna was determined to be 0.705 (nominal: 0.826) mg test item/L. The observed 5 % mortality at this concentration level is in the control data range. The 48h-LOEC was 1.569 (nominal 1.818) mg test item/L. While the 48h-EC50 was calculated to be 1.59 mg/L (nominal: 1.75 mg/L); the 48h-EC0 of the test item was 0.288 mg/L (nominal 0.376 mg/L) and the 48h-EC100 was determined to be 9.4 (nominal 8.8) mg test item/L.

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

For read-across justification please refer to IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

1.59 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.705 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

For read-across justification please refer to IUCLID section 13.

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.56 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.102 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

from 2008-02-05 to 2008-02-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

The study is regarded as reliable without restrctions because it was conducted in accordance with GLP regulation and guideline.

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Sampling method:
For determination of the test item concentration, samples were taken from the testing concentrations and the control at the start and at the end of the test.
- Sample storage conditions before analysis:
The samples were analysed directly after sampling. The samples will be kept stored in deep-freezer up to the date of the final report.

Vehicle:

no

Details on test solutions:

A stock solution with a concentration of 80 mg/L was prepared with test item and ISO Medium by means of ultrasonic treatment two days before the start of the study (on day -2). The test solutions were prepared from this stock solution by the appropriate dilution with ISO Medium on day 0 as given in the table below. The stock solution and all test solutions were prepared without the use of organic solvents. The nominal concentration of the test item was confirmed by HPLC (nominal 80 mg/L, measured 81 mg/L).
(In the main study four replicates were used for each test concentration and for the untreated control.)
Start of the test: immediately after preparation of the test solutions Daphnids were introduced (< 15 minutes).

Concentration (mg/L) Stock solution (mL) ISO Medium (mL) Total volume (mL)
nominal measured
Control - 300 300.0
0.625 0.540 2.4 300 302.4
1.25 1.102 4.8 300 304.8
2.5 2.47 9.7 300 309.7
5.0 5.21 20.0 300 320.0
10.0 10.2 43.0 300 343.0

Control: In the control, test water was used without addition of the test item.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species: Daphnia magna (Straus) not first brood progeny
Age at Test Start: Less than 24 hours old
Sex: Female
Origin: National Institute of Public Health, Hungary
Breeding Conditions: The Daphnia were bred in the laboratories of LAB Research Ltd. under similar temperature and light conditions as in the test, and in reconstituted water of a similar quality regarding to pH, components of the main ions and total hardness as the test water used in the test. The test animals were fed daily with a few mL of centrifuged algae suspension (Pseudokirchneriella subcapitata).
Acclimatisation: Brood Daphnids were maintained in ISO medium under test conditions prior to the start of the test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

169.6 mg/L

Test temperature:

20.6 - 21.0 °C

pH:

7.45 to 8.17

Dissolved oxygen:

6.4 mg/L or higher

Nominal and measured concentrations:

nominal: 0.625; 1.25; 2.5; 5.0 and 10.0 mg/L
measured: 0.540; 1.102; 2.47; 5.21 and 10.2 mg/L

Details on test conditions:

Test Units
Type and Size: Glass beakers of 40 mL test medium
Identification: Each test unit was uniquely identified with study code, treatment and replicate number.

Test Conditions
Test Environment: Climate chamber
Temperature: 18 – 22 °C with a maximum deviation of ± 1 °C
Light period: 16 h light : 8 h dark, light intensity: 912 lux
Recording: Test conditions were measured with suitable instruments and documented in the raw data.

Test water
Reconstituted Water: ISO medium (during the holding period and during the test).
The hardness (CaCO3) of water was: 169.6 mg/L 1). 1) Determined by National Public Health and Medical Officer Service. Date of sampling: 05 February 2008; Results: 07 February 2008, The documentation is archived in the raw data notebook.
The ISO medium (according to OECD 202) was prepared by adding 25 ml each of the following stock solutions to deionised water.
Substance
CaCl2 x 2 H2O 11.76 g/L
Batch No.: KBR5022501
Supplier: REANAL
Expiry date: May 2009
MgSO4 x 7H2O 4.93 g/L
Batch No.: A895286
Supplier: MERCK
Expiry date: 31 Aug. 2012
NaHCO3 2.59 g/L
Batch No.: 829880
Supplier: REANAL
Expiry date: Dec. 2007
KCl 0.23 g/L
Batch No.: PP/2007/09359/0
Supplier: REANAL
Expiry date: 31 May 2010

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.54 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

3.56 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 10.2 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

1.25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

2.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

10.2 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

1.102 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

1.102 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

2.47 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Analytical Results
The analytically determined mean test item concentrations in the analysed test media varied in the range from 101 % to 107 % at the start and from 72 % to 105 % at the end of the study in comparison to the nominal value.
Biological Results and Conclusion
Results after 48 hours:
The 48h-NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), respectively the 48h-EC0 of Reaction products of MXDA with bisphenol A diglycidylether (BADGE) to Daphnia magna was determined to be 1.102 (nominal: 1.25) mg test item/L. The 48h-LOEC was 2.47 (nominal 2.5) mg test item/L, while the 48h-EC50 was calculated to be 3.56 (nominal: 3.54) mg test item/L and the 48h-EC100 was determined to be 10.2 (nominal 10.0) mg test item/L.

Results with reference substance (positive control):

For the evaluation of the quality of the Daphnia clone and the experimental conditions, potassium dichromate is tested at least twice a year to demonstrate satisfactory test conditions.
The date of the last study (Study Code: 07/541-023DA) with reference item was: 20 - 21 November 2007. The results of the reference study are the followings:
The 24h-EC50 value: 1.86 mg /L
95 % confidence limits: 1.50 – 2.21 mg /L

In the test with the reference item the results met the expected toxicity of the POTASSIUM DICHROMATE (the 24h EC50 value of the potassium dichromate should be within the range 0.6 mg/L to 2.1 mg/L).The results are based on the nominal concentrations.

Reported statistics and error estimates:

The EC50 values of the test item and their confidence limits were calculated using Probit analysis. The analysis was done using the statistical software program “TOXSTAT 3.5”.
The EC0, the NOEC, the LOEC and the EC100 were determined directly from the raw data.

Validity criteria fulfilled:

yes

Conclusions:

In the 48-hour static acute toxicity test to Daphnia magna with Reaction products of MXDA with bisphenol A diglycidylether (BADGE) the EC50 value was determined to be 3.56 mg/L (Probit analysis). The overall NOEC was determined to be 1.102 mg/L.

Executive summary:

The purpose of this study was to evaluate the influence of the test item Reaction products of MXDA with bisphenol A diglycidylether (BADGE) on the mobility respectively survival of Daphnia magna. Young Daphnia were exposed in a static test to the test item for 48 hours, added to test water at a range of concentrations.

0.625; 1.25; 2.5; 5.0 and 10.0 mg/L nominal test item concentrations and a control were tested in the main study. The corresponding mean measured test item concentrations were 0.540; 1.102; 2.47; 5.21 and 10.2 mg test item/L.

The test item concentration of the test solutions was measured by HPLC-UV method at the start and at the end of the test.

The analytically determined test item concentrations in the analysed test media varied in the range from 101 % to 107 % at the start and from 72 % to 105 % at the end of the study in comparison to the nominal value. The deviation of the measured concentrations from the nominal values was higher than 20 % at the lowest two concentration levels at the end of the test.

Therefore, all reported biological results are related to the geometric mean measured concentrations of the test item at the start and at the end of the test calculated by EXCEL Software Program.

The 48h-NOEC (highest concentration tested without toxic effects after the exposure period of 48 hours), respectively the 48h-EC0 of Reaction products of MXDA with bisphenol A diglycidylether (BADGE) to Daphnia magna was determined to be 1.102 (nominal: 1.25) mg test item/L. The 48h-LOEC was 2.47 (nominal 2.5) mg test item/L, while the 48h-EC50 was calculated to be 3.56 (nominal: 3.54) mg test item/L and the 48h-EC100 was determined to be 10.2 (nominal 10.0) mg test item/L.

Description of key information

Reaction Product of Bisphenol A diglycidylether (BADGE) with a mixture of IPDA and MXDA was not tested for short-term ecotoxicological properties. Instead, a read-across approach was applied for short-term toxicity to aquatic invertebrates using data of symmetrical epoxy amine adducts Reaction products of IPDA with bisphenol A diglycidylether (BADGE) and Reaction products of MXDA with bisphenol A diglycidylether (BADGE), as these substances were considered to show similar ecotoxicological properties. Both symmetrical epoxy amine adducts were tested for short-term toxicity to invertebrates according to EU method C.2 and OECD 202. Determined worst-case toxicity values for the asymmetrical epoxy amine adduct Reaction Product of Bisphenol A diglycidylether (BADGE) with IPDA and MXDA were a 48-h EC50 of 1.59 mg/L (mean measured value) and 48-h NOEC of 0.705 mg/L.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1.59 mg/L

Additional information

The asymmetrical epoxy amine adduct Reaction Product of Bisphenol A diglycidylether (BADGE) with IPDA and MXDA was not tested for short-term ecotoxicological properties. Instead, a read-across approach was applied to assess short-term toxicity to aquatic invertebrates, using data from the symmetrical epoxy amine adducts Reaction products of IPDA with bisphenol A diglycidylether (BADGE) and Reaction products of MXDA with bisphenol A diglycidylether (BADGE), as these substances were considered to show similar ecotoxicological properties.

In both these studies, young Daphnia were exposed in a static test for 48 hours. Stock solutions were prepared with test item and ISO Medium by means of ultrasonic treatment two days before the start of the studies (on day -2). Test solutions were prepared from stock solutions by appropriate dilution with ISO Medium on day 0. The stock solutions and all test solutions were prepared without the use of organic solvents. The concentrations of the test item were confirmed by HPLC nominal: 0.171; 0.376; 0.826; 1.818; 4.0 and 8.8 mg/L; measured: 0.136; 0.288; 0.705; 1.569; 4.6 and 9.4 mg/L and nominal: 0.625; 1.25; 2.5; 5.0 and 10.0 mg/L; measured: 0.540; 1.102; 2.47; 5.21 and 10.2 mg/L for Reaction products of IPDA with bisphenol A diglycidylether (BADGE) and Reaction products of MXDA with bisphenol A diglycidylether (BADGE), respectively.

The 48-hour static acute toxicity tests with Daphnia magna, according to EU method C.2 and OECD 202, revealed a 48-h EC50 values of 1.59 mg/L and 3.59 mg/L (nominal: 1.75 mg/L and 3.54 mg/L) and the 48-h NOEC values were determined to be 0.705 mg/L and 1.102 mg/L (nominal: 0.826 mg/L and 1.25 mg/L), respectively for Reaction products of IPDA with bisphenol A diglycidylether (BADGE) and Reaction products of MXDA with bisphenol A diglycidylether (BADGE).

Toxicity values from the most toxic symmetrical adduct were used for read-across approach to the asymmetrical adduct Reaction product of Bisphenol A diglycidylether (BADGE) with IPDA and MXDA, representing the worst case scenario. The short-term toxicity to aquatic invertebrates was a 48-h EC50 of 1.59 mg/L and a 48-h NOEC of 0.705 mg/L.