Choline chloride

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-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: see 'Remark'

Remarks:

Experiment is reported in the freely available IUCLID 4 substance dataset from ESIS. No study report was handed out. The experiment was conducted according to OECD 211 and GLP compliance without detailed information. Available data indicate that the study was well-performed.

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes

Remarks:

No further details available.

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material: No surrogate or analogue material was used.

Analytical monitoring:

yes

Details on sampling:

The test concentrations were measured for both renewal and old test solution at the start of the experiment and after 1h, 10th, 11th, 20th and 21th day.

Vehicle:

no

Details on test solutions:

The test substance was diluted with dilution water. The test substance was stored in the freezer. The stability of the chemical was confirmed by IR absorption spectrum. Under the stock condition, IR spectrum of the test substance at the end of the test was the same at the start.

Test organisms (species):

Daphnia magna

Details on test organisms:

- Age: < 24 h old
- Supplier / Source: Test organisms were obtained from the National Institute of Environmental Studies (Japan)
- Pretreatment: Parental daphnids were acclimated for 20 days on test condition before testing.
- Feed: Chlorella vulgaris, 0.15 mg carbon/day/individual
- Mothers of test individuals were selected from batches which were not observed death individuals and any resting-eggs and male daphnids.

Test type:

other: static renewal

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

No post exposure observation period described.

Hardness:

23 - 37 mg/L (as CaCO3)

Test temperature:

20.3 - 21.0 °C

pH:

7.0 - 7.7

Dissolved oxygen:

6.7 - 8.1 mgO2/L

Salinity:

Not applicable.

Nominal and measured concentrations:

0, 3.2, 10, 32 and 100 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: Erlenmeyer flask
- Material, size, fill volume: glass, 200 mL, 80 mL fill volume
- Aeration: none
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: prepared from dechlorinated industrial water (drinkable water grade). This water was aerated after residual chlorine removal by activated carbon treatment
- pH: 7.3
- Total hardness (as CaCO3): 37 mg/L

OTHER TEST CONDITIONS
- Photoperiod: 16 h dark, 8 h light
- Feeding: 0.15 - 0.18 mg carbon/day/individual (Chlorella vulgaris, green algae)

RANGE-FINDING STUDY
None reported.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 95.5 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

other: parental mortality

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

58.9 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: 95 % CL: 37.7 - 83.7 mg/L

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

30.2 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

95.5 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

No further details available.

Results with reference substance (positive control):

EC50(48h): 0.079 mg/L (based on mobility)

Reported statistics and error estimates:

- LC50 with corresponding 95 % CL: cannot be calculated
- EC50 with corresponding 95 % CL: calculated by Probit method
- NOEC and LOEC: The cumulative number of juveniles produced per adult in control and test concentration after 21 d was tested by Dunnett multiple comparison method
- Method of calculating mean measured concentrations: Time-weighted Mean

Table 1. Measured concentrations

Nominal concentration [mg/L]	0h New	1h Old	10d New	11d Old	20d New	21d Old
0.00	N.D.	N.D.	N.D.	N.D.	N.D.	N.D.
3.20	3.10	3.10	3.30	2.50	3.20	1.90
10.00	9.70	9.70	10.30	8.10	9.90	6.60
32.00	33.50	32.80	34.20	25.70	32.10	23.70
100.00	104.00	101.00	104.00	87.00	101.00	77.30
New: freshly prepared test solution. Old: test solutions 24 h after freshly prepared.

Table 2. Percent of nominal concentration

Nominal concentration [mg/L]	0h New	1h Old	10d New	11d Old	20d New	21d Old
0.00	--	--	--	--	--	--
3.20	97 %	97 %	103 %	78 %	100 %	59 %
10.00	97 %	97 %	103 %	81 %	99 %	66 %
32.00	105 %	103 %	107 %	80 %	100 %	74 %
100.00	104 %	101 %	104 %	87 %	101 %	77 %
New: freshly prepared test solution. Old: test solutions 24 h after freshly prepared.

Table 3. Cumulative number of died parental Daphnia

Nominal concentration [mg/L]	Cumulative Number of Died Parental Daphnia after 21 days [Mortality rate in %]
0.00	0 (0)
3.20	0 (0)
10.00	2 (20)
32.00	0 (0)
100.00	0 (0)

Table 4. Time to first brood production

Nominal concentration [mg/L]	Time to First Brood Production [Days]
0.00	8.3
3.20	7.9
10.00	8.9
32.00	8.3
100.00	11.2

Table 5. Cumulative numbers of juvenile produced per adult

Nominal Concentration [mg/L]	Mean cumulative numbers of juveniles produced per adult for 21 days
	0 - 6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
0.00	0 - 0	1.0	11.8	12.0	14.0	36.5	36.6	43.0	66.4	66.4	74.2	105.9	106.1	111.5	140.0	140.0
3.20	0 - 0	4.8	10.6	11.0	24.0	37.6	37.6	50.1	67.2	67.2	82.4	106.5	106.5	116.9	164.0	146.0
10.00	0 - 0	0.1	8.3	9.0	9.90	32.6	32.8	39.8	63.5	63.5	70.3	103.8	103.9	109.5	142.0	142.0
32.00	0 - 0	3.7	8.8	8.8	13.3	30.6	30.7	41.4	58.3	58.3	72.4	94.1	94.1	115.4	136.1	136.1
100.00	0 - 0	0.0	0.0	0.3	0.80	2.70	4.10	4.10	5.60	7.5	8.9	9.5	11.8	11.8	11.8	11.8

Table 6. Cumulative numbers of juvenile produced per adult alive for 21 days

Nominal concentration [mg/L] (Measured concentration [mg/L])
Vessel No.	0.00	3.20 (2.83)	10.00 (9.00)	32.00 (30.20)	100.00 (95.50)
1	162	150	D	140	13
2	151	136	147	147	12
3	152	143	125	114	5
4	142	150	D	168	12
5	150	154	139	125	11
6	136	124	151	125	12
7	128	152	152	142	10
8	118	140	129	129	17
9	121	162	140	149	11
10	140	149	153	122	15
Mean	140.0	146.0	142.0	136.1	11.8
S.D.	14.4	10.7	10.7	16.1	3.2
Inhibition ratio [%]	-	-4.3	-1.4	2.8	91.6
Significant difference	-	N.S.	N.S.	N.S.	*
D: No calculation performed since parental Daphnia was dead during the 21-days testing period N.S.: Indicate a no-significant difference by Dunnett multiple comparison procedure *: Indicate a significant difference by Dunnett multiple comparison procedure

Validity criteria fulfilled:

yes

Remarks:

Scientific criteria for acute toxicity testing to aquatic invertebrates seemed to be fulfilled.

Conclusions:

The study is reported in the freely available IUCLID 4 substance dataset from ESIS. The experiment was conducted according to OECD 211 with GLP compliance. Available data indicate that the study was well-performed. As result, the NOEC(21d) was determined as 30.2 mg/L, based on time-weighted mean of measured concentrations.

Executive summary:

A long-term experiment was conducted with the aquatic invertebrate Daphnia magna in order to determine the impact of Choline chloride on the reproduction rate during an exposure duration of 21 days (MOE Japan, 1999). The study was conducted according to OECD 211 and under GLP compliance. The nominal test concentrations were reported as 0, 3.2, 10, 32 and 100 mg/L, whereby the measured concentrations (based on time-weighted mean of measured concentrations) were as followed: 0, 2.83, 9.00, 30.2 and 95.5 mg/L. 10 replicates per concentration and 1 individual per replicate were implemented. The stability of the test substance was confirmed by IR absorption spectrum. A photoperiod of 16 h light and 8 h dark was produced. The green algae Chlorella vulgaris served as food during the exposure duration. Potassium dichromate was used as reference substance, resulting in an EC50(48h) of 0.079, based on mobility). The effect data for choline chloride, based on reproduction, were reported as followed: LC50(21d) > 95.50 mg/L (parental mortality), EC50(21d): 58.9 mg/L (95 % CL: 37.7 - 83.7 mg/L), NOEC(21d): 30.2 mg/L and LOEC(21d): 95.5 mg/L. The results are based on the time-weighted mean of measured concentrations.

Description of key information

Daphnia magna_OECD 211_time-weighted mean of measured concentrations: LC50(21d) > 95.50 mg/L (parental mortality), EC50(21d): 58.9 mg/L (95 % CL: 37.7 - 83.7 mg/L), NOEC(21d): 30.2 mg/L, LOEC(21d): 95.5 mg/L

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Effect concentration:

30.2 mg/L

Additional information

A long-term experiment was conducted with the aquatic invertebrate Daphnia magna in order to determine the impact of Choline chloride on the reproduction rate during an exposure duration of 21 days (MOE Japan, 1999). The study was conducted according to OECD 211 and under GLP compliance. The nominal test concentrations were reported as 0, 3.2, 10, 32 and 100 mg/L, whereby the measured concentrations (based on time-weighted mean of measured concentrations) were as followed: 0, 2.83, 9.00, 30.2 and 95.5 mg/L. 10 replicates per concentration and 1 individual per replicate were implemented. The stability of the test substance was confirmed by IR absorption spectrum. A photoperiod of 16 h light and 8 h dark was produced. The green algae Chlorella vulgaris served as food during the exposure duration. Potassium dichromate was used as reference substance, resulting in an EC50(48h) of 0.079, based on mobility). The effect data for choline chloride, based on reproduction, were reported as followed: LC50(21d) > 95.50 mg/L (parental mortality), EC50(21d): 58.9 mg/L (95 % CL: 37.7 - 83.7 mg/L), NOEC(21d): 30.2 mg/L and LOEC(21d): 95.5 mg/L. The results are based on the time-weighted mean of measured concentrations.

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