Trimethylammonium chloride

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:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions (e.g. non-GLP, no analytical verification of test concentrations)

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Version / remarks:

Directive 79/831/EEC, Annex V, Part C2

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: Institut National de Recherche Chimique Appliquée, France; kept and bred since 1978 at BASF AG, Laboratory for Environmental Analytics and Ecology
- Age at study initiation (mean and range, SD): 2-24 h
- Method of breeding:
- Water conditions:
- Total hardness: 2.70 ± 0.5 mmol/L
- Ks up to pH 4.3: 0.80 ± 0.1 mmol/L
- Molar ratio: about 4:1
- Dissolved oxygen: > 2 mg/L
- Feeding: brewer's yeast at water exchange; washed green algae culture once daily
- Illumination: 16 h light, 8 h dark
- Feeding during test
- Food type: no details available
- Amount: no details available
- Frequency: no details available

ACCLIMATION
- Acclimation period: mp details available
- Acclimation conditions (same as test or not): no details available
- Type and amount of food: no details available, assumed to be identical as stated above: brewer's yeast at water excahnge, wahsed green algae culture once daily
- Feeding frequency: no details available, assued to be identical as stated above: once daily
- Health during acclimation (any mortality observed): not specified

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

2.52 mmol/L

Test temperature:

19.05 °C

pH:

7.95

Dissolved oxygen:

saturated (aerated till saturation with oil-free air)

Nominal and measured concentrations:

Nominal test concentrations: 0 (control), 7.81, 15.6, 31.2, 62.5, 125, 250, and 500 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size: 20-mL glass tubes with flat bottom
- Test volume: 10 mL
- Aeration: none
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: 2 ml/animal

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted freshwater
- tap water, filtered (6 µm) and dechlorinated (activated carbon)
- addition of H2SO4 to reduce Ks pH 4.3
- addition of destilled water to reduce total hardness
- aeration with oil-free air until saturated with oxygen
- storage of prepared water for 24 h for stabilisation
- Total hardness: 2.70 ± 0.50 mmol/L
- Acid capacity (Ks) to pH 4.3: 0.80 ± 0.1 mmol/L
- Ca/Mg ratio: ca. 4:1
- Na/K ratio: ca. 10:1
- Conductivity: 550-650 µS/cm
- pH 7.7-8.3
- Culture medium different from test medium: no

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 h light, 8 h dark
- Light intensity: ca. 5 µE/m²/s at 400-750 nm
- Temperature: 20-22 °C

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mobility: 0, 3, 6, 24, and 48 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2

Reference substance (positive control):

no

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

125 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

259.13 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

- Immobility (number of immobile animals after x hours):

Concentration (mg/L)		0 h		3 h		6 h		24 h		48 h
0 (control)		0		0		0		1		1
7.81		0		0		0		0		0
15.6		0		0		0		1		1
31.2		0		0		0		0		0
62.5		0		0		0		0		0
125		0		0		0		1		2
250		0		0		0		0		6
500		0		0		0		9		20

- pH and dissolved oxygen content:

Concentration	pH		dissolved oxygen (mg/L)
(mg/L)	0 h	48 h	0 h	48 h
0 (control)	7.91	7.67	8.40	8.1
7.81	7.80	7.82	8.90	8.1
15.6	7.81	7.73	8.60	7.6
31.2	7.81	7.79	8.30	8.2
62.5	7.80	7.84	8.70	8.2
125	7.77	7.84	8.70	8.4
250	7.73	7.82	8.70	8.2
500	7.69	7.83	8.70	8.3

Specifications of test water (measured at test start)

- Total hardness: 2.52 mmol/L

- Acid capacity (Ks) to pH 4.3: 0.85 mmol/L

- Conductivity: 625 µS/cm

- pH 7.95

Validity criteria fulfilled:

yes

Conclusions:

An EC50 (48 h) of 259.13 mg/L was determined - based on nominal test concentration and mobility as evaluated effect.

Executive summary:

The short-term toxicity to aquatic invertebrates of the test substance trimethylamine chlorohydrate (CAS 593 -81 -7) has been investigated according to EU Directive 79/831/EEC, Annex V, part C. GLP compliance for this static freshwater test is not given. As test organism Daphnia magna was used. The total exposure duration amounts to 48 hours. Nominal substance concentrations (mg/L) of 0 (control), 7.81, 15.60, 31.20, 62.50, 125, 250 and 500 mg/L were tested (no analytical verification). Four replicates per concentration and for the control have been used, respectively. Each test vessel (flat bottom glass tubes) contained 5 daphnids. A photoperiod of 16:8 hours (day-night period) was chosen.

As final results and based on mobility, an EC0 (48 h) of 125 mg/L, an EC 50 (48 h) of 259.13 mg/L and an EC 100 (48 h) of 500 mg/L were determined. The results are based on nominal test concentrations.

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: Guideline study with acceptable restrictions (e.g. non-GLP, no analytical verification of test concentrations)

Qualifier:

according to guideline

Guideline:

other: EU Directive 79/831/EEC, Annex V, part C

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

Preparation of test media:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The test medium of the highest test concentration, 500 mg/L (nominal) was prepared. Following this, serial dilutions of this test medium were prepared in order to obtain the desired test concentrations. 10 mL of the respective dilutions were added to the test vessels.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: Institute National de Recherche Chimique Appliquee, France. Cultured in the laboratories of BASF since 1978.
- Age at study initiation (mean and range, SD): 2-24 h
- Feeding during test: none

ACCLIMATION
- Type and amount of food: Whilst changing water Brewer's yeast was given, also 1 x daily grown Green algae culture

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Post exposure observation period:

no data

Hardness:

2.70 ± 0.5 mmol/L

Test temperature:

19.85 °C

pH:

start: 8.22 - 10.49
48 h: 8.07 - 9.33

Dissolved oxygen:

start: 8.51 - 8.62
48 h: 7.94 - 8.12

Salinity:

no data

Nominal and measured concentrations:

Nominal concentrations (mg/L):
control, 15.62, 31.20, 62.50, 125.00, 250.00, 500.00

Details on test conditions:

TEST SYSTEM
- Test vessel: flat bottom glass tubes
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water; pretreatment steps: 6 µm- and charcoal-filtration; H2SO4 was added to reduce alkalinity up to pH 4.3; distilled water was added to reduce water-hardness; water was aerated (oil-free air) unil saturated with oxygen; water was stored for at least 24 h for stabilization.
- Ca/mg ratio: 4:1

OTHER TEST CONDITIONS
- Photoperiod: 16:8 day-night period
- Light intensity: diffuse light (5 microeinstein/m*m*s) at 400 - 750 nm wave length

Reference substance (positive control):

not specified

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

62.5 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

250 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

139.95 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Remarks on result:

other: 95 % confidence limits: 123.1 - 154.6 mg/L

Table 1. Cumulative immobilization of Daphnia after x hours [%]

Nominal test concentration (mg/L)	3 h	6 h	24 h	48 h
Control	0	0	0	0
15.62	0	0	0	0
31.2	0	0	0	0
62.5	0	0	0	0
125	0	0	10	35
250	50	20	100	100
500	100	100	100	100

Remark in the report: toxicity at 250 mg/L and 500 mg/L may be due to the high pH (up to 10.49)

Table 2: pH values after 0 & 48 hours
Conc. [mg/L]	0 h	48 h
0	8.22	8.07
15.62	8.75	8.11
31.20	9.13	8.17
62.50	9.50	8.26
125.00	9.86	8.51
250.00	10.19	8.97
500.00	10.49	9.33

Validity criteria fulfilled:

yes

Conclusions:

An EC50 (48 h) of 139.95 mg/L was determined - based on nominal test concentration and mobility as evaluated effect.

Executive summary:

The short-term toxicity to aquatic invertebrates of the read-across substance trimethylamine (CAS 75-50-3) has been investigated according to EU Directive 79/831/EEC, Annex V, part C. GLP compliance for this static freshwater test is not given. As test organism, Daphnia magna was used. The total exposure duration amounts to 48 hours. Nominal substance concentrations (mg/L) of 0 (control), 15.62, 31.20, 62.50, 125, 250 and 500 mg/L were tested (no analytical verification). Four replicates per concentration and for the control have been used, respectively. Each test vessel (flat bottom glass tubes) contained 5 daphnids. A photoperiod of 16:8 hours (day-night period) was chosen. Observed toxicity at 250 mg/L and 500 mg/L may be due to the high pH (up to 10.49).

As final results and based on mobility, an EC0 (48 h) of 62.5 mg/L, an EC 50 (48 h) of 139.95 mg/L and an EC 100 (48 h) of 250 mg/L were determined. The results are based on nominal test concentrations.

In addition to that, a screening tests was performed in order to determine the evaporation of trimethylamine closely following the test conditions.

Based on the measured total carbon values it can be stated, that there is no significant loss of the test substance.

Description of key information

Trimethylamine chlorohydrate_EU Directive 79/831/EEC, Annex V, part C_Daphnia magna_EC50 (48 h) = 259.13 mg/L

Trimethylamine (read-across)_EU Directive 79/831/EEC, Annex V, part C_Daphnia magna_EC50 (48 h) = 139.95 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

259.13 mg/L

Additional information

The short-term toxicity to aquatic invertebrates of the test substance Trimethylamine chlorohydrate (CAS 593 -81 -7) has been investigated according to EU Directive 79/831/EEC, Annex V, part C (Key study 1, 1987). GLP compliance for this static freshwater test is not given. As test organism Daphnia magna was used. The total exposure duration amounts to 48 hours. Nominal substance concentrations (mg/L) of 0 (control), 7.81, 15.60, 31.20, 62.50, 125, 250 and 500 mg/L were tested (no analytical verification). Four replicates per concentration and for the control have been used, respectively. Each test vessel (flat bottom glass tubes) contained 5 daphnids. A photoperiod of 16:8 hours (day-night period) was chosen.

As final results and based on mobility, an EC0 (48 h) of 125 mg/L, an EC 50 (48 h) of 259.13 mg/L and an EC 100 (48 h) of 500 mg/L were determined. The results are based on nominal test concentrations.

In addition, the short-term toxicity to aquatic invertebrates of the read-across substance Trimethylamine (CAS 75-50-3) has been investigated according to EU Directive 79/831/EEC, Annex V, part C (Supporting study 1, 1988; 2002). GLP compliance for this static freshwater test is not given. As test organism, Daphnia magna was used. The total exposure duration amounts to 48 hours. Nominal substance concentrations (mg/L) of 0 (control), 15.62, 31.20, 62.50, 125, 250 and 500 mg/L were tested (no analytical verification). Four replicates per concentration and for the control have been used, respectively. Each test vessel (flat bottom glass tubes) contained 5 daphnids. A photoperiod of 16:8 hours (day-night period) was chosen. Observed toxicity at 250 mg/L and 500 mg/L may be due to the high pH (up to 10.49).

As final results and based on mobility, an EC0 (48 h) of 62.5 mg/L, an EC 50 (48 h) of 139.95 mg/L and an EC 100 (48 h) of 250 mg/L were determined. The results are based on nominal test concentrations.

In addition to that, a screening test was performed in order to determine the evaporation of trimethylamine closely following the test conditions.

Based on the measured total carbon values it can be stated, that there is no significant loss of Trimethylamine.

Trimethylamine and Trimethylammonium chloride belong to the group of tertiary aliphatic amines with a sp³-hybridized nitrogen with three methyl groups (common structure / functional group). The solvation of both Trimethylamine and Trimethylammonium chloride in water results in solutions of the Trimethylammonium cation (common "breakdown product"). Both respective counterions are naturally and ubiquitous occurring ions and are also to a certain extent required for the maintenance of various body functions. Besides the influence on the pH value of an aqueous solution (OH-), they do not bear a relevant intrinsic property, allowing one in general to focus on the Trimethylammonium cation. Toxicity of the respective counterion is not applicable (Cl-) in the aqueous environment, a slightly aggravating effect is expected for the hydroxyl anion (OH-). So primarily the trimethylammonium cation has to be evaluated. It is believed to be act and to be metabolised by the same mechanisms by microorganisms and by other classes of living organisms. Therefore both substances are expected to follow the same pattern of toxicity to fish. For the detailed procedure of the read-across principle and justifications, please refer to the analogue approach justification depicted below and the separate Read-Across Statement (Chemservice S.A., 2015).