2,4,6-tris(dimethylaminomethyl)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

Study period:

2018 -03-08 to 2018-10-12

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Concentration: 100 mg/L
Samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test.
sampling volume : 5 mL
Storage: Samples were stored in a freezer (<= -15 °C) until analysis at the analytical laboratory of the test facility.

Vehicle:

no

Details on test solutions:

A limit test with a nominal concentration of 100 mg/L based on the results of a preceeding range-finding test.
Controls: Test medium without test item or other additives.

Test organisms (species):

Daphnia magna

Details on test organisms:

Species Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
Source In-house laboratory culture with a known history.
Reason for selection This test species has been selected as an internationally accepted invertebrate species.
Validity of batch 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.
Characteristics Daphnia, less than 24 hours old, from parental daphnids of more than two weeks old.
Breeding
Start of each batch Approximately 250 newborn daphnids, i.e. less than 3 days old, were placed into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures 4 weeks
Renewal of the cultures After 7 days of cultivation, half of the medium twice a week.
Temperature of medium 18-22°C
Feeding Daily, a suspension of fresh water algae.
Culture medium M7, as prescribed by Dr. Elendt-Schneider
(Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).
Feeding during test No feeding

Test type:

static

Water media type:

other: Adjusted ISO medium, formulated using RO-water (tap-water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands)

Limit test:

yes

Total exposure duration:

48 h

Hardness:

180 mg CaC03 per liter

Test temperature:

between 20 and 22°C

pH:

Start: 8.0 ; End : 8.3 in control
Start: 8.0 ; End : 8.1 in test concentration

Dissolved oxygen:

Start: 8.3 mg/l; End: 9.3 (control)
Start: 8.3 mg/l; End: 9.3 (test contrentration)

Nominal and measured concentrations:

Nominal: 100 mg/L
Measured concentrations: 92 -94 % of nominal throughout the exposure period.

Details on test conditions:

- Test vessel: 60 mL all-glass
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 50 mL
- Aeration: no aeration during the test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 5 per vessel containing 50 mL of test solution
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: adjusted ISO-Medium prepared by adding following salts(analytical grade) to tap water purified by Reverse Osmosis (RO-water, GEON Waterbehandeling, Berkel-Enschot, The Netherlands): CaCl2.2H2O 211.5 mg/L
MgSO4.7H2O 88.8 mg/L
NaHCO3 46.7 mg/L
KCl 4.2 mg/L
- Total organic carbon: 180 mg CaCO3 per liter
- Alkalinity: pH between 6 and 9
- Culture medium different from test medium: yes (M7-medium was used for the culture and adjusted ISO-Medium for the test)
- Intervals of water quality measurement: pH and dissolved oxygen were measured at t=0 h and t=48 h. The temperature was measured continuously in a temperature control vessel, beginning at the start of the test
OTHER TEST CONDITIONS
- Adjustment of pH: yes. The pH of the test solution was adjusted using a 1M HCl solution (Merck, Darmstadt, Germany) from 9.6 to 7.7 in the limit test.
- Photoperiod: A daily photoperiod of 16 hours
- Light intensity: not specified
EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
VEHICLE CONTROL PERFORMED: no
RANGE-FINDING STUDY
- Test concentrations: 0.10, 1.0 and 10 mg/L in the combined range-finding test.
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Key result

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: No mortality observed: Test item not toxic.

Key result

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: No toxicity observed.

Details on results:

No EC50 could be calculated because the test item proved to be non-toxic (EC50 > regulatory limit concentration).

Results with reference substance (positive control):

Results with reference substance valid: yes
- Mortality/immobility: yes
- LC50: 0.53 mg/L
- Other: Study according to OECD guideline No.202 (2004) in a static procedure.
Under the conditions of the test, the 48h-LC50 for Daphnia magna exposed to Potassium dichromate was 0.59 mg/L (95 % confidence interval between 0.53 and 0.72 mg/L).

pH-values and dissolved oxygen concentrations (mg/L) during the limit test

Nominal concentration (mg/L)	start (t= 0h)		End (t= 48 h)
	pH	O2	pH	O2
Control	8.0	8.3	8.3	9.3
100	8.0	8.3	8.1	9.3

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the test item proved to have no effect on the mobility of Daphnia magna up to the regulatory limit analytically confirmed nominal concentration of 100 mg/L.

Executive summary:

The ability of the test item to generate acute toxic effects on the mobility of Daphnia magna was examined according to the OECD guideline No. 202, (2004).

A limit test was performed based on the results of a preceding range-finding test. Twenty daphnids per group (5 per replicate, quadruplicate) were exposed in a static procedure to an untreated control and to nominally 100 mg/L, during an exposure period of 48 hours. Samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test.

Measured concentrations were 92‑94% of nominal throughout the exposure period. Based on these results, the effect parameters were expressed as analytically confirmed nominal concentrations.

No immobility was observed in the control and at any of the test concentrations throughout the exposure period.

In conclusion, the test item proved to have no effect on the mobility of Daphnia magna up to the regulatory limit nominal concentration of 100 mg/L. The test item is not toxic to daphnids.

The validity criteria according to the OECD202 was fulfilled.

Description of key information

A short-term toxicity testing on aquatic invertebrates with standard organisms according to standardized guideline was required within a CCH decision.

Accordingly, the ability of the test item to generate acute toxic effects on the mobility of Daphnia magna was examined according to the OECD guideline No. 202, (2004).

A limit test was performed based on the results of a preceding range-finding test. Twenty daphnids per group (5 per replicate, quadruplicate) were exposed in a static procedure to an untreated control and to nominally 100 mg/L, during an exposure period of 48 hours. Samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test. Measured concentrations were 92‑94% of nominal throughout the exposure period. Based on these results, the effect parameters were expressed as analytically confirmed nominal concentrations. No immobility was observed in the control and at any of the test concentrations throughout the exposure period.

In conclusion, the test item proved to have no effect on the mobility of Daphnia magna up to the regulatory limit nominal concentration of 100 mg/L. The test item is not toxic to daphnids.

The validity criteria according to the OECD202 was fulfilled.

Additional information

No EC50/LC50 could be calculated because the test item proved to be non-toxic in the limit test conducted at an analytically confirmed nominal concentration of 100 mg/L.