Reaction mass of 2,6-Bis[(dimethylamino)methyl]-4-(1-{3-[(dimethylamino)methyl]-4-hydroxyphenyl}-1-methylethyl)phenol and 4-(1-{3,5-Bis[(dimethylamino)methyl]-4-hydroxyphenyl}-1-methylethyl)-2,6-bis[(dimethylamino)methyl]phenol

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:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Remarks:

HPLC/MS/MS

Details on sampling:

Control: at 48 hours
Test concentration/s: 0.40, 0.88, 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/L at 0 and 48 hours

Vehicle:

no

Remarks:

mortar and pestle, ultrasonic bath, magnetic stirrer

Details on test solutions:

Pre-treatment of test item and preparation of test item concentrations
- test item was pulverized with a mortar and a pestle.
- a stock solution was prepared to give the desired series of test concentrations
- 100.6 mg of the test item were added to 1 litre of dilution water, treated for 1 h in an ultrasonic bath and stirred for 24 h on a magnetic stirrer
- pH was measured to be 9.2 and was adjusted to pH 8.7
- to produce the different test item concentrations appropriate amounts of the stock solution were diluted with dilution water to a volume of 500 mL
- 50 mL of the prepared solutions were taken per replicate and 0.5 mL of dilution water was given to all replicates resulting in the final nominal concentrations

Test organisms (species):

Daphnia magna

Details on test organisms:

- Name: Daphnia magna STRAUS, parthenogenetic females
- Source: Strain of Bundesgesundheitsamt Berlin
- Maintenance and Acclimatisation: A population of parthenogenetic females of synchronized age structure has been maintained for more than 20 years in the test facility under constant temperature conditions (20 +/- 2 °C) at a 16 : 8 hour light-dark photoperiod (light intensity:
< 20 µE x m-2 x s-1). The culture water (so-called 'M4 medium') was partly renewed once a week. The Daphnia were exclusively fed unicellular green algae (Desmodesmus subspicatus) 'ad libitum'. Mortalities of parent Daphnia during the culture period were recorded daily. The neonates were separated from their parent Daphnia by filtration prior to the acute test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

14.4 °dH (= 257 mg/L CaCO3)

Test temperature:

20.0 - 20.6 °C

pH:

7.9 - 8.8

Dissolved oxygen:

8.8 - 9.1 mg/L

Nominal and measured concentrations:

nominal [mg/L]: 0.40, 0.88, 1.9, 4.3, 9.4, 20.7, 45.5, 100
Measured (geometric mean; Mannichbase 1, 2 and 3) [mg/L]: 0.46, 0.93, 1.67, 3.20, 5.91, 19.39, 35.74, 83.49
Effective concentrations of the sum of Mannichbase 1, 2 and 3 ranged from 68.7 % to 121.2 % of nominal values at 0 hours and from 57.6 % to109.9 % of nominal values at 48 hours.

Details on test conditions:

Exposure conditions
- Test vessels: 100 mL glass beakers covered with watch glasses holding 5 neonates in 50 mL of test medium
- Experimental design: 8 test concentrations plus 1 control, 5 neonates per vessel, 4 replicates per concentration/control,
- no feeding during the exposure period
- static system
- Method of initiation: neonates were placed in prepared media
- Photoperiod: 16 h light : 8 h dark
- Temperature of incubation unit: 19.6 to 20.2 °C
- Aeration: none
- Test item concentration/s: 0.40, 0.88, 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/L (nominal)
- Method of administration: stock solution (100 mg/L)
- Medium renewal: none
- Duration of exposure: 48 hours
- Criteria of effects: item-induced alteration of the normal mobility behaviour and the loss of locomotory actions of the neonates, observed at 24 and 48 hours

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

16.4 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

The results are expressed in terms of geometric mean measured concentrations as the sum of Mannichbase 1 (2x substituted), 2 (3x substituted) and 3 (4x substituted).
Effective concentrations of the sum of Mannichbase 1, 2 and 3 ranged from 68.7 % to 121.2 % of nominal values at 0 hours and from 57.6 % to109.9 % of nominal values at 48 hours.

Reported statistics and error estimates:

The EC 50 was calculated by probit analysis using the statistics programme ToxRatPro Version 2.10 (released 2010-09-10).

Validity criteria fulfilled:

yes

Remarks:

immobilisation and other abnormalities in the controls did not exceed 10 % by the end of the test; dissolved oxygen concentration remained above 3 mg/L throughout the exposure period

Conclusions:

After48 h an EC50 of 16.4 mg/L was determined for HPP 12879-1 towards Daphnia under exposure conditions.

Executive summary:

A study was performed to assess the acute toxicity of HPP 12879-1 to Daphnia magna STRAUS under static conditions.

It was conducted in accordance with the Council Regulation (EC) No 440/2008, Method C.2 ‘Acute toxicity for Daphnia’ (2008) which is equivalent to OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test' (adopted April 13, 2004).

A range finding test preceded the main test. It provided information about the range of concentrations which were used in the main test. The following nominal concentrations of the test item were tested in the range finding test: 1, 10 and 100 mg/L.

The Daphnia were exposed to a range of concentrations, nominally 0.40, 0.88, 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/L of HPP 12879-1 dissolved in dilution water for a period of 48 hours. Auxiliaries used to prepare the test media were an ultrasonic bath and a magnetic stirrer. The test was conducted as a static test with the test medium unchanged throughout the duration of the test. During the test a temperature range of 18 - 22 °C was maintained in the test vessels, with a maximum temperature fluctuation of +/- 1 °C in each individual test. The temperature, the pH and the oxygen values were measured at the beginning and end of the test.

Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. The EC 50 was calculated by probit analysis using the statistics programme ToxRatPro Version 2.10 (released 2010-09-10). Additionally any abnormal behaviour or appearance of the Daphnia was reported. The following value was determined 48h-EC50 = 16.4 mg/L.

The results are expressed in terms of geometric mean measured concentrations as the sum of Mannichbase 1 (2x substituted), 2 (3x substituted) and 3 (4x substituted).Effective concentrations of the sum of Mannichbase 1, 2 and 3 ranged from 68.7 % to 121.2 % of nominal values at 0 hours and from 57.6 % to109.9 % of nominal values at 48 hours.

Description of key information

A study was performed to assess the acute toxicity of HPP 12879-1 to Daphnia magna STRAUS under static conditions.

It was conducted in accordance with the Council Regulation (EC) No 440/2008, Method C.2 ‘Acute toxicity for Daphnia’ (2008) which is equivalent to OECD Guideline for Testing of Chemicals No. 202 'Daphnia sp., Acute Immobilisation Test' (adopted April 13, 2004).

The Daphnia were exposed to a range of concentrations, nominally 0.40, 0.88, 1.9, 4.3, 9.4, 20.7, 45.5 and 100 mg/L of HPP 12879-1 dissolved in dilution water for a period of 48 hours. Auxiliaries used to prepare the test media were an ultrasonic bath and a magnetic stirrer. The test was conducted as a static test with the test medium unchanged throughout the duration of the test. During the test a temperature range of 18 - 22 °C was maintained in the test vessels, with a maximum temperature fluctuation of +/- 1 °C in each individual test. The temperature, the pH and the oxygen values were measured at the beginning and end of the test.

Observations were made on the swimming ability and the immobilisation rate, respectively, after 24 and 48 hours of exposure. The EC 50 was calculated by probit analysis using the statistics programme ToxRatPro Version 2.10 (released 2010-09-10). Additionally any abnormal behaviour or appearance of the Daphnia was reported. The following value was determined 48h-EC50 = 16.4 mg/L.

The results are expressed in terms of geometric mean measured concentrations as the sum of Mannichbase 1 (2x substituted), 2 (3x substituted) and 3 (4x substituted).Effective concentrations of the sum of Mannichbase 1, 2 and 3 ranged from 68.7 % to 121.2 % of nominal values at 0 hours and from 57.6 % to109.9 % of nominal values at 48 hours.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

16.4 mg/L