3-aminomethyl-3,5,5-trimethylcyclohexylamine

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

1984

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

1992

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

The analytical verification of the test item concentrations was carried out after 0 and 48 hours. For stability control an additional series of the same concentrations was prepared in separate vessels without test organisms

Vehicle:

no

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION
- Concentration: 1.04 g/l; 0.52 g of the test item were equilibrated directly in 500 ml synthetic fresh water.

DILUTION WATER
- Source: Synthetic:
CaCl2 x 2 H2O: 294 mg/l
MgSO4 x 6 H2O: 114 mg/l
NaHCO3: 65 mg/l
KCl: 6 mg/l
- Ca/Mg ratio: 4:1
- Na/K ratio: 10:1

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISMS
- Strain: Daphnia magna Straus, clone 5
- Source/supplier: received from Bayer AG in 1991, further bred inhouse
- Breeding method: in 1 l beakers with M4 medium, water renewal each 2-3 days, isolation of juveniles for further breeding each ca. 4 weeks
- Age: < 24 hours
- Feeding: Desmodesmus subspicatus, as much as consumed
- Pretreatment: Filtration of adults 24 h prior to testing
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

14 degree German hardness = 250 mg CaCO3/l

Test temperature:

20.22 - 20.28 (mean 20.25) degree C

pH:

Study start: 7.6 (control); 7.9; 8.2; 8.5; 8.9; 9.2; 9.6 (highest conc.)

Dissolved oxygen:

48 hours: 7.8 (control); 7.9; 7.9; 7.9; 8.1; 8.0; 8.2 (highest conc.) mg O2/l

Nominal and measured concentrations:

Nominal: 0.0; 2.1; 4.2; 8.3; 16.6; 33.2; 66.4 mg/l
Measured: - ; 1.9; 3.9; 7.7; 15.5; 32.1; 66.3 mg/l (48 hours)

Details on test conditions:

- Exposure vessel type: 10 ml round-bottom test tubes
- Number of replicates, individuals per replicate: 4 replicates with 5 individuals each
- Intensity of irradiation: dark
- Photoperiod: no
- Aeration: no
- Control group: 1 blank control simultaneously and 2 reference substance control

TEST PARAMETER: immobilisation

Reference substance (positive control):

yes

Remarks:

Potassium dichromate, CAS RN 7778-50-9

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

66.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

23 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 17-31 mg/L (95% confidence limits)

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

8.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: 18-40 mg/L (95% confidence limits).

Details on results:

The EC50-values were calculated by probit analysis according to Cavalli-Sforza (1972)

Results with reference substance (positive control):

- Concentrations: 1.0; 2.0 mg/l
- Results: 40; 100 % immobilization
- tested every 3 months

Reported statistics and error estimates:

- 95 % confidence interval of EC50:
18 - 40 mg/l (24 hours); EC50 = 27 mg/l
17 - 31 mg/l (48 hours); EC50 = 23 mg/l

RESULTS: EXPOSED
- Concentration / response table:

conc. in mg/l nominal	0.0	2.1	4.2	8.3	16.6	33.2	66.4
% immobile (24 hours)	0	0	0	15	30	40	100
% immobile (48 hours)	0	0	0	10	30	65	100

Validity criteria fulfilled:

not specified

Conclusions:

The EC50 (48 hours) was determined as 23 mg/L indicating that the test substance may be harmful to aquatic invertebrates.

Executive summary:

The acute toxicity of 3 -aminomethyl-3,5,5 -trimethylcyclohexylamine to Daphnia magna was studied under static conditions over a period of 48 hours according to OECD TGD 202 and EU-method C.2. Six concentrations ranging from 2.1 to 66.4 mg/L were tested. Immobilisation was observed. The following endpoint was derived:

EC50 (48 h): 23 mg/L; CL: 17-31 mg/L

The study was classified as "reliable without restrictions".

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test procedure generally in accordance with national standard methods with acceptable restrictions (e.g. no controls)

Qualifier:

according to guideline

Guideline:

other: DIN 38412, part 11

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION
- Concentration: 1 g/l

Test organisms (species):

Daphnia magna

Details on test organisms:

- Strain: Daphnia magna, Huels
- Source/supplier: Huels AG (inhouse)
- Breeding method: in 1 l jars with dechlorinated drinking water, water renewal every 2-3 days, isolation of juveniles for further breeding every ca. 4 weeks
- Age: < 24 hours
- Feeding: Chlorella vulgaris, as much as consumed
- Pretreatment: Filtration of adults 24 h prior to testing
- Feeding during test: no

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Test temperature:

20 +/- 1 degree C

Nominal and measured concentrations:

Nominal concentrations: 12; 18; 25; 35; 50; 70; 100 mg/l

Details on test conditions:

- Exposure vessel type: 25 ml graduated cylinder
- 7 test concentrations + 2 reference substance concentrations (no blank)
- Number of replicates, individuals per replicate: 4 replicates with 5 individuals each
- Intensity of irradiation: dark
- Photoperiod: no (dark)
- Aeration: no

Reference substance (positive control):

yes

Remarks:

potassium dichromate, CAS RN 7778-50-9

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

25 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

70 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

44 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 35-50 mg/L (95% Confidence limits).

Details on results:

- Concentration / response table:
12; 18; 25; 35; 50; 70; 100 mg/l
0; 0; 0; 35; 60; 100; 100 % immobile

Results with reference substance (positive control):

- Concentrations: 0.9; 1.9 mg/l
- Results: 10; 65 % immobilization

Reported statistics and error estimates:

- 95 % confidence interval of LC50: 35-50 mg/l

Validity criteria fulfilled:

not specified

Conclusions:

The 24h-EC50 was determined as 44 mg/L indicating that the test substance may be harmful to aquatic invertebrates.

Executive summary:

The acute toxicity of 3-aminomethyl-3,5,5-trimethylcyclohexylamine to Daphnia magna was studied under static conditions over a period of 24 hours according to DIN 38412, part 11. Seven concentrations ranging from 12 to 100 mg/L plus control were tested. Immobilisation was observed. The following endpoint was derived:

EC50(24 h): 44 mg/L; CL: 35-50 mg/L

The study was classified as "reliable with restrictions".

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Scientifically acceptable publication

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

no

Analytical monitoring:

not specified

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: An appropriate amount of test substance was weighed into the test medium establishing a stock solution of a nominal concentration of 500 mg/l. In order to facilitate the solution, the mixture was ultrasonicated for 5 min. followed by magnetic stirring for 22 hours. After separation of test medium and possible remaining particles or droplets for 2 hours, aliquots were sampled from the mid-fraction of the stock solution for preparation of the test concentrations.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: isolated from Lake Langedam, Birkerod, Denmark in 1979 and has been cultured at the VKI (Water Quality Institute) since then.
- Age at study initiation (mean and range, SD): < 24 h
- Method of breeding: The animals are cultured in water from Lake Bradebaek (Denmark) at 20 °C fed three times every day with Raphidocelis subcapitata and a supply of yeast cells once or twice every week.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

250 ±25 mg/l CaCO3

Test temperature:

20 °C +/- 1 °C

pH:

7.8 +/- 0.1

Dissolved oxygen:

% Saturation after
24 h
99 - 100 %
48 h
96 - 97 %

Nominal and measured concentrations:

Nominal concentrations (mg/l)
control
5
10
20
40
80
160

Details on test conditions:

TEST SYSTEM
- Test vessel: glass vessels (capacity 250 ml)
- Type (delete if not applicable): covered with plastic sheets
- Fill volume: 125 ml
- Renewal rate of test solution (frequency/flow rate): After 24 hours, the test animals were transferred to new test vessels prepared as specified
under "Details on test solutions".
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 6

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Freshly produced 150-medium was used in the tests. The medium was prepared from deionised water, and salts were added to the water according to the standard procedure (IS0 International Standard 6341)

OTHER TEST CONDITIONS
- Adjustment of pH: to 7.8 +/- 0.1
- Photoperiod: 16:8 day-night regime

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mobility

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

20.7 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

37.4 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

other: EC90

Effect conc.:

54 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

8.33 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

17.4 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

other: EC90

Effect conc.:

26.4 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Conclusions:

The 24 h EC50 based on mobility was 37,4 mg/L, the 48 h EC50 baased on mobility was 17,4 mg/ L, indicating that the test substance may be harmful to aquatic invertebrates.

Executive summary:

The acute toxicity of the test substance to Daphnia magna was studied over a period of 48 hours according to OECD 202. Six concentrations ranging from 5 to 160 mg/L were tested. Immobilisation was observed. The following endpoint was derived:

EC50 (48 h): 17,4 mg/ L

Reference 4

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Principles of method if other than guideline:

Method: other: see Test Conditions

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

not specified

Details on test solutions:

The test solutions were prepared from concentrated stock solutions and naural seawater. Where necessary, an organic solvent (preferably Dimethylsulfoxide) was used to prepare the stock solution (as 24 compounds were tested not clear, if used for this particular test).

DILUTION WATER (=seawater)
- Source: Eastern Scheldt (NL), prepared by sand filtration, filtration over activated charcoal and 0.2 um millipore filter
- pH: about 8
- Control group: natural seawater

Test organisms (species):

other aquatic crustacea: Chaetogammarus marinus

Details on test organisms:

TEST ORGANISMS
- Breeding method: Readily grown in seawater aquarium systems with shelter for concealment
- Age: Young gammarids, about 5 mm long
- Feeding: Fucus spec. - Feeding during test: yes (Fucus or Tetramin); reason: to prevent cannibalism

Test type:

semi-static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

15 +/- 1 degree C

pH:

8.0 (control); 8.5; 8.8; 9.1; 9.5; 10.0; 10.2; 10.2 (highest conc.)

Dissolved oxygen:

almost saturated for the whole test

Salinity:

28 o/oo

Nominal and measured concentrations:

Nominal: 0; 32; 56; 100; 180; 320; 560; 1000 mg/l

Details on test conditions:

TEST CONDITIONS
- Renewal of test solution: once a day
- Aeration: no
- Exposure vessel type: 1 l glass beakers with 1 l test solution, covered with a watch glass
- Number of replicates, individuals per replicate: 2 beakers with 10 animals each for each concentration
- Adjustment of pH: no
- solvent control, if solvent is used

TEST PARAMETER: mortality; counting and removal of dead animals daily accompanied by visual inspection of survivors

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality and behaviour compared to the control

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

572 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 505-648 mg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

388 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 229-444 mg/L

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

362 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 318-412 mg/L

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

324 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 286-366 mg/L

Reported statistics and error estimates:

LC50-values and error estimates (confidence limits)
24 hours: 572 (505-648) mg/l
48 hours: 388 (339-444) mg/l
72 hours: 362 (318-412) mg/l
96 hours: 324 (286-366) mg/l

Validity criteria fulfilled:

not specified

Conclusions:

The 96 hour-LC50 was determined as 324 mg/l.

Executive summary:

The aquatic toxicity of 3-aminomethyl-3,5,5-trimethylcyclohexylamine was tested in the marine invertebrate Chaetogammarus

marinus under semistatic conditions. Five concentrations ranging from 32 to 1000 mg/L were tested. The test organisms were observerd daily for mortality and abnormal behaviour / symptoms. The 96 hour-LC50 was determined as 324 mg/l.

 

The study was assessed as "valid without restrictions".

Description of key information

Cited from SIAR for SIAM 18 (Paris, France, April 20–23, 2004):
"The acute toxicity of 3-aminomethyl-3,5,5-trimethylcyclohexylamine to Daphnia magna was determined in a static test conducted according to OECD 202 (I) (1984). After 48 h of exposure, the EC50 was calculated to 23 mg/l (Infracor GmbH, 2002b). In a test according to DIN 38412, part 11 a nominal EC50 (24 h) of 44 mg/l was reported (Hüls AG, 1996a). The aquatic toxicity of 3-aminomethyl-3,5,5-trimethylcyclohexylamine was also tested in the marine invertebrate Chaetogammarus marinus. The 96 hour-EC50 determined in this semistatic test is 324 mg/l (Adema, 1982). However, despite of good test performance and documentation, the result with this non-standard organism may at present only serve as an indication that the sensitivity of marine invertebrates towards the test substance is probably not higher than that of freshwater organisms."

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

EC50

Effect concentration:

23 mg/L

Marine water invertebrates

Marine water invertebrates

Dose descriptor:

EC50

Effect concentration:

324 mg/L

Additional information

For the freshwater environment, the study on Daphnia magna conducted by Infracor GmbH, 2002 is considered as key study because it was conducted under GLP according to accepted scientific guidelines and shows the lowest toxicity endpoint that are analytically verified. For the marine environment, the study reported by Adema, 1982 is taken into account, but it has to be noted that a non-standard test organism was used and that the test design was not in line with any accepted scientific guideline.