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

Study period:

4 JAN 1999 - 30 JUL 1999

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes

Remarks:

The study complied with the Environment Agency's GLP regulations.

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:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Eluate: Choline chloride at a concentration of 1000 mg/L was prepared as a stock solution with dilution water, and the prescribed volume was placed in a 1 L flask and diluted to a constant volume with dilution water to make a test solution. The test solution was prepared in a 1 L female flask and diluted with dilution water to a constant volume.
- Other relevant information: The test solution was analysed at a frequency of 3 times during the exposure period (6 times in total before and after the water change) for all concentration groups (but one test container per test container). The test water was analysed by HPLC. When analysing the test water. The standard solution was measured for each sample measurement and quantified from its peak height. 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.
- Method of breeding: Adult females of sufficient size to be healthy and visually healthy with juveniles were selected from those undergoing passaging and transferred to a separately prepared beaker. The next day, the produced juveniles were divided into another beaker. The larvae were reared under the following conditions. The larvae were removed at least twice a week once they had matured and started to lay young.
Breeding water: Dilution water
Breeding density: 15 head/L Breeding water
Water temperature: 20 +- 1 °C
Illumination: Room light, 16 h light/8 h dark
- 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:

semi-static

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 - 7.6

Dissolved oxygen:

6.7 - 8.1 mgO2/L

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: 80 mL in 200mL flask with stopper
- Aeration: semi-stopped water system (water change for 24 hours), Dechlorinated water (industrial water (chlorinated and of drinking water grade) from the Nishiki Plant of Kureha Chemical Industry Co. grade) was treated with activated carbon to remove residual chlorine, etc., and aerated with air.
- No. of organisms per vessel: 10 animals/concentration (1 animal per series).
- No. of vessels per concentration (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)

After measuring the water temperature, dissolved oxygen concentration (DO), pH and hardness of the test solution, a glass pipette was The test sample Daphnia magna was introduced using a glass pipette, at which point the exposure was initiated. At that time, the The total amount of breeding water in the pipette was kept within 1 % of the test solution volume. Thereafter, at each water change Daphnia daphnia were transferred to a new test solution after each water change and reared until day 21. During the exposure period, Daphnia magna were fed at a constant rate every day. A fixed amount of food was fed daily during the exposure period.

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

> 95.5 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Remarks on result:

other: Parental Daphnia magna

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

58.9 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

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.

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

95.5 mg/L

Nominal / measured:

meas. (TWA)

Conc. based on:

test mat.

Details on results:

- Mortality of control: The mortality rate of the parent Daphnia in the control area was 0 % at the end of the exposure. in nominal concentration 10 % on day 14 and 21 mortality 10 and 20 % observed. This meets the criteria of less than 20 %, which was a condition for the study to be established. The criteria for the study to be valid, i.e. less than or equal to 20 %, were met. No dormant eggs were observed in the control and in each test concentration.

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 experiment was conducted according to OECD 211 with GLP compliance. As result, the NOEC(21d) was determined as 30.2 mg/L, based on time-weighted mean of measured concentrations, LOEC was derived as 95.5 mg/L. EC50 value for the 21-day exposure was 58.9 mg/L, LC50 of parental daphnia magna was determined as > 95.5 mg/L.

Executive summary:

A long-term experiment was conducted with the aquatic invertebrate Daphnia magna in order to determine the impact of test item 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. 10 replicates per concentration and 1 individual per replicate were implemented. 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.

Description of key information

Daphnia magna_OECD 211: 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 test item 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. 10 replicates per concentration and 1 individual per replicate were implemented. 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.