Diethylammonium chloride

Toxicity to aquatic algae and cyanobacteria

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Please refer to the attached analogue justification provided in IUCLID Section 13.

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

20 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: cell growth based on fluorimetric units

Remarks on result:

other: Source: Calamari et al., 1980

Duration:

96 h

Dose descriptor:

EC0

Effect conc.:

2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: cell growth based on fluorimetric units

Remarks on result:

other: Source: Calamari et al., 1980

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

50.86 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: Source: 6.1.5-1

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

34.27 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: Source: 6.1.5-1

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

56 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: Source: Van Leeuwen et al., 1985

Reference 2

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: EPA, algal assay with Selenastram capricornutum applied to toxicity tests as proposed by Chiaudani and Vighi, Mitt. Internat. Veretn. Limnol., 21,316 (1978).

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

- Sampling method: the concentrations of the substance in water tests were determined at the beginning of the experiments and frequently during, or at the end of the tests.

Vehicle:

no

Test organisms (species):

Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

7 d

Details on test conditions:

EFFECT PARAMETERS MEASURED: growth was evaluated by measuring in vivo the fluorimetric units (according to C.J. Lorenzen, Deep Sea Res., 13, 223, 1968) at 48, 72, 96 h and 7 d.

Reference substance (positive control):

not specified

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

20 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: cell growth based on fluorimetric units

Remarks on result:

other: slope of the effect concentration curve was 0.2

Duration:

96 h

Dose descriptor:

EC0

Effect conc.:

2 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

other: cell growth based on fluorimetric units

Remarks on result:

other: slope of the effect concentration curve was 0.2

Other biological results: EC100 (96 h) = 80 mg/L

Validity criteria fulfilled:

not specified

Reference 3

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 - 14 April 2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

1984

Deviations:

no

GLP compliance:

yes

Remarks:

according to the GLP standard by the Japanese Ministry of Environment

Specific details on test material used for the study:

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: at room temperature in the dark

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all test concentrations were sampled and measured at the beginning and at the end of the test

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: 100 mg/L test substance was diluted in test medium. This stock solution was filtered and further diluted with test medium up to the respective concentrations.

Test organisms (species):

Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: green algae
- Age of inoculum (at test initiation): 3 d

ACCLIMATION
- Acclimation period: 3 d
- Culturing media and conditions (same as test or not): same as test
- Any deformed or abnormal cells observed: not observed

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test temperature:

22.0 - 23.1 °C

pH:

7.4 - 7.2 (control), 7.4 - 10.0 (test item)

Nominal and measured concentrations:

0 (control), 4.8, 8.6, 15.4, 27.8, 50.0, 90.0 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: 300 mL conical flask
- Type: closed
- Material, size, headspace, fill volume: glass, 100 mL
- Aeration: no
- Initial cells density: 1 x 10^4 cells/mL (verified by counting chamber)
- Control end cells density: 194.6 x 10^4 cells/mL (mean)
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 3

GROWTH MEDIUM
- Standard medium used: yes

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: according to guideline
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH was measured at the beginning and at the end of the test

OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: continuously
- Light intensity and quality: 4000 - 5000 lux

EFFECT PARAMETERS MEASURED
- Determination of cell concentrations: counting chamber

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.8
- Range finding study was performed but the result was not reported in the test report

Reference substance (positive control):

yes

Remarks:

potassium dichromate

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

50.86 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: values calculated in retrospect with ToxRatPro v. 3.2.1 based on the original cell densities stated in the study report

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

34.27 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: values calculated in retrospect with ToxRatPro v. 3.2.1 based on the original cell densities stated in the study report

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

48.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

15.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Details on results:

- Exponential growth in the control (for algal test): yes, increase by a factor of 195
- Observation of abnormalities (for algal test): no abnormalities observed

Results with reference substance (positive control):

- Results with reference substance valid? yes
- EC50: 0.52 mg/L

Reported statistics and error estimates:

Probit, Barlett, ANOVA, Dunnet

Table 1: Measured concentrations of diethylamine

Nominal concentration (mg/L)	Measured concentration (mg/L)
	0 h	% of nominal	72 h	% of nominal
Control	< 0.04	-	< 0.04	-
4.8	4.0	83.3	2.1	43.8
8.6	7.4	86.0	4.1	47.7
15.4	14.8	96.1	7.5	48.7
27.8	26.8	96.4	13.3	47.8
50.0	45.3	90.6	24.0	48.0
90.0	87.3	97.0	46.0	51.1

 

 

Table 2: Cell Density

Nominal Concentration (mg/L)		Cell Density (x 10^4 cells/mL)
		0 h	24 h	48 h	72 h
Control	1	1	5.8	38.7	193.3
	2	1	5.3	41.8	198.2
	3	1	6.1	39.6	192.4
	mean		5.733	40.033	194.633
4.8	1	1	4.9	34.4	179.7
	2	1	4.6	38.2	190.5
	3	1	5.5	39.2	202.8
	mean	1	5	37.267	191
8.6	1	1	5.1	36.5	198.6
	2	1	5.4	38.2	189.6
	3	1	5.7	37.8	184.4
	mean	1	5.4	37.5	190.867
15.4	1	1	5.4	36.3	200
	2	1	5.2	35.4	188.4
	3	1	4.9	35.3	182.4
	mean	1	5.167	35.667	190.267
27.8	1	1	4.7	39.6	152.2
	2	1	5.6	38.7	147.1
	3	1	4.9	37.8	146.1
	mean	1	5.067	38.7	148.467
50	1	1	4.1	28.4	129.4
	2	1	3.8	29.9	132.9
	3	1	3.8	30.1	124.4
	mean	1	3.9	29.467	128.9
90	1	1	1.5	3.1	4
	2	1	1.5	2.6	2.8
	3	1	1.5	3.1	3.8
	mean	1	1.5	2.933	3.533

Table 3: % Inhibition of growth rate at 72 h as calculated in retrospect according to the current OECD 201 standards by ToxRatPro v. 3.2.1 using the orginal cell densities from the study report.

Geom. mean measured concentrations (mg/L)	% Inhibition at 72 h
Control	-
2.89	0.38
5.51	0.38
10.54	0.44
18.88	5.14
32.97	7.82
63.37	76.28

Validity criteria fulfilled:

yes

Conclusions:

The 72h (growth rate) LC50 of the test substance ammounts to 54 mg/L and the NOEC of 11 mg/L was reported when tested under the conditions of this assay (OECD TG 202).

Reference 4

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study without detailed documentation

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

GLP compliance:

not specified

Analytical monitoring:

not specified

Details on sampling:

- Sampling method: samples were taken with syringes

Vehicle:

not specified

Test organisms (species):

Chlorella pyrenoidosa

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

20±1°C

Details on test conditions:

TEST SYSTEM
- Test vessel: 200 mL Erlenmayer flasks, 100 mL test volume
- Type: stoppered with a cotton plug
- Initial cells density: 10E8 cells/L
- No. of organisms per vessel: 10E5 cells
- other: slight agitation on a mechanical shaker
- No. of vessels per concentration (replicates): 3

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: The composition of the medium used is given in Van Leeuwen and Maas, 1985

OTHER TEST CONDITIONS
- Sterile test conditions: no
- Photoperiod: continuously
- Light intensity and quality: fluorescent light at 7.5 W/m2

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: ZBI Coulter Counting chamber

Duration:

96 h

Dose descriptor:

EC50

Effect conc.:

56 mg/L

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

growth rate

Validity criteria fulfilled:

not specified

Conclusions:

The 96h (growth rate) LC50 of diethylamine ammounts to 56 mg/L when tested under the conditions of this assay (OECD TG 201).

Description of key information

ErC50 (96 h) = 20 mg/L, ErC0 (96 h) = 2 mg/L (meas., EPA algal assay as proposed by Chiaudani and Vighi, 1978) based on read across from CAS 109-89-7

Key value for chemical safety assessment

EC50 for freshwater algae:

20 mg/L

EC10 or NOEC for freshwater algae:

2 mg/L

Additional information

Experimental data on the toxicity of Diethylammonium chloride (CAS 660-68-4) towards algae are not available. Therefore appropriate read across data from the analogues substance N-ethylethanamine (CAS 109-89-7) were used for the environmental assessment.

The toxicity of N-ethylethanamine (CAS 109-89-7) to the algae species Selenastrum capricornutum was tested in the study by Calamari et al., 1980, according to EPA, algal assay with Selenastrum capricornutum as proposed by Chiaudani and Vighi (1978). The concentrations of the substance in the test media were determined at the beginning of the experiments and frequently during, or at the end of the test by gaschromatography. Growth was determined by measuring in vivo the fluorimetric units at 48, 72, 96 h and 7 d. The study resulted in an ErC50 (96 h) of 20 mg/L and an ErC0 (96 h) of 2 mg/L (meas.). These results were regarded as key information since they represent the most sensitive effect values of the trophic level.

The toxicity of the read across substance N-ethylethanamine (CAS 109-89-7) to algae was tested by a further study according to OECD 201 and GLP standards (reference: 6.1.5). In a 72 h study, the test organism Pseudokirchneriella subcapitata was exposed to nominal test item concentrations of 0 (control), 4.8, 8.6, 15.4, 27.8, 50.0 and 90.0 mg/L (nominal). The stability of the tested substance in the medium was analytically monitored by HPLC. 43.8 – 51.1% of the substance was recovered after 72 hours at the test end. Since no growth rate effect values from 0 -72 h were reported in the study report, a calculation of the ErC50 (72 h) and ErC10 (72 h) was performed in retrospect with ToxRatPro v.3.2.1 using the original cell numbers stated in the report. The calculation resulted in an ErC50 (72 h) of 50.86 mg/L and an ErC10 (72 h) of 34.27 mg/L (both geom. mean meas.).

Supporting information is available from a publication by Van Leeuwen et al. (1985). The toxicity of N-ethylethanamine (CAS 109-89-7) to algae was tested according to the OECD guideline 201. Chlorella pyrenoidosa was exposed to the read across substance for 96 h. The study resulted in an ErC50 (96 h) of 56 mg/L (nominal).