m-phenylenebis(methylamine)

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:

No data

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to internationally accepted guidelines. OECD Guideline 211. No deviations

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

Sampling was conducted every 24 hours

Vehicle:

no

Details on test solutions:

Nominal concentrations: Control, 0.150, 0.470, 1.50, 4.70, 15.0 mg/L

Test organisms (species):

Daphnia magna

Details on test organisms:

No data

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

No data

Hardness:

195-255 mg/L of calcium carbonate - see Table 11 (attached)

Test temperature:

20 ± 1 degrees Centigrade - see Table 8 (attached)

pH:

pH 7.4 - 8.8 - see Table 10 (attached)

Dissolved oxygen:

7.5 - 8.8 mg/L - see Table 9 (attached)

Salinity:

No data

Nominal and measured concentrations:

Nominal concentrations: control, 0.150, 0.470, 1.50, 4.70, 15.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: closed with the surace of the water covered with a teflon sheet
- Fill volume: 80 mL
- Renewal rate of test solution: Every 24 hours
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of individuals per dose level: 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Elendt M4 medium - see Table A-1 (attached)

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8 hours darkness
- Light intensity: 800 lux

EFFECT PARAMETERS MEASURED: Mortality and reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

4.7 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Duration:

21 d

Dose descriptor:

EC50

Effect conc.:

8.4 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: Unable to calculate 95% confidence limit

Duration:

21 d

Dose descriptor:

LC50

Effect conc.:

6.77 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: 95% confidence limit 1.5 - 15.0 mg/L

Details on results:

All of the measured concentrations of the test material were within ± 20 % of nominal concentrations, therefore, the nominal concentrations were used for calculation of effect concentration. Representative chromatograms are attached.

No environmental factors which might affect the reliability of the study results were seen.

The mortality of parental Daphnia in the control group was 0 % at the end of the exposure period, and this result fulfilled the validation criteria for the study (20 % or less). The mortality in the highest concentration group was 100 % at the end of the exposure period.

First brood production by parent Daphnia in the control group took place on Day 8 after the start of exposure, which was considered to be within the normal range. All parental Daphnia in the highest concentration group died before first brood production.

Mean cumulative numbers of juveniles produced per one parent Daphnia during 21 days in the control group were 108 animals, and this fulfilled the validation criteria of the study (60 animals). All parental Daphnia in the highest concentration group died before first brood production.

Generation of dormant broods was not observed in the control group or any test concentration group during the exposure period.

The temperatures of the test solutions were 20 ± 1 degrees Centigrade in all concentration groups, and dissolved oxygen concentrations were 60 % or more of the saturated concentration (at 20 degrees Centigrade: 8.8 mg/L). Both of these values fulfilled the study standards. The pH values were within the appropriate range (6.0 ~ 9.0 with a variation of 1.5) for raising Daphnia. In addition, the hardness was considered to be within the appropriate range (approximately 250 mg/L).

The results of test material measurement in the test solutions during the exposure period are shown in Tables 1 -1 and 1 -2 (below):

Table 1-1 - Measured Concentrations of the Test Substance in Test Water during a 21 -day Exposure Period (Daphnia Reproduction Inhibition Test under Semi-Static Test Conditions)

Nominal Concentration (mg/L)	Measured Concentration (mg/L)							TWM *^1(mg/L)	% of Nominal
	Date	0	1	7	8	14	15
		New	Old	New	Old	New	Old
Control		<0.007	<0.007	<0.007	<0.007	<0.007	<0.007	-	-
0.150		0.172	0.150	0.163	0.150	0.157	0.148	0.157	105
0.470		0.542	0.484	0.536	0.498	0.523	0.497	0.513	109
1.50		1.70	1.55	1.71	1.60	1.60	1.56	1.62	108
4.70		4.84	4.71	4.85	4.78	4.81	4.71	4.78	102
15.0		15.4	15.3	*	*	*	*	15.3	102
New	Freshly prepared test solution
Old	Old test solution before renewal
*1	Time-weighted mean measured concentration during 21 days
*	No measurement was made because all parental Daphnia were dead

Table 1 -2 - Measured Concentrations as a Percentage of Nominal

Nominal Concentration (mg/L)	Measured Concentration as a Percentage of Nominal
	Date	0	1	7	8	14	15
		New	Old	New	Old	New	Old
0.150		115	100	109	100	105	99
0.470		115	103	114	106	111	106
1.50		113	103	114	107	107	104
4.70		103	100	103	102	102	100
15.0		103	102	*	*	*	*
New	Freshly prepared test solution
Old	Old solution before renewal
*1	Time-weighted mean measured concentration during 21 days
*	No measurement was made because all parental Daphnia were dead

Cumulative mortality of parental Daphnia during the exposure period are shown in Tables 2 -1 and 2 -2 (below) and Figure 1 (attached):

Table 2 -1 - Cumulative Number of Dead Parental Daphnia

 

Days	Nominal concentration
	Control	0.150 mg/L	0.470 mg/L	1.50 mg/L	4.70 mg/L	15.0 mg/L
0	0	0	0	0	0	0
1	0	0	0	0	1	1
2	0	0	0	0	1	1
3	0	0	0	0	1	5
4	0	0	0	0	1	9
5	0	0	0	0	2	10
6	0	0	0	0	2	10
7	0	0	0	0	2	10
8	0	0	0	0	2	10
9	0	0	0	0	2	10
10	0	0	0	0	2	10
11	0	0	0	0	2	10
12	0	0	0	0	2	10
13	0	0	0	0	2	10
14	0	1	0	0	2	10
15	0	1	0	0	2	10
16	0	1	0	0	2	10
17	0	1	0	0	2	10
18	0	1	0	0	2	10
19	0	1	0	0	2	10
20	0	1	0	0	2	10
21	0	1	0	0	2	10

 

Table 2 -2 - Mortality (%) of Parental Daphnia

 

Nominal concentration	Days
	1	2	4	7	14	21
Control	0	0	0	0	0	0
0.150 mg/L	0	0	0	0	10	10
0.470 mg/L	0	0	0	0	0	0
1.50 mg/L	0	0	0	0	0	0
4.70 mg/L	10	10	10	20	20	20
15.0 mg/L	10	10	90	100	100	100

 

The days to the first brood production by parental Daphnia are shown in Table 3 (below) for each concentration:

Table 3 - Time (Days) to First Brood Production

Vessel Number	Nominal Concentration mg/L
	Control	0.150	0.470	1.50	4.70	15.0
1	8	8	8	8	8	*
2	8	8	8	10	9	*
3	8	8	8	8	9	*
4	8	8	8	8	*	*
5	8	8	8	8	*	*
6	8	8	8	8	8	*
7	8	8	8	8	8	*
8	8	8	8	8	10	*
9	8	8	8	8	8	*
10	8	9	8	8	9	*
Min	8	8	8	8	8	*
Max	8	9	8	10	10	*
*	The parental Daphnia died before first brood production

Mean cumulative numbers of juveniles produced per adult alive for the exposure period in each concentration are shown in Table 4 (below) and Figure 2 (attached):

Table 4 - Mean Cumulative Numbers of Juveniles Produced per Adult Alive for 21 Days (ΣFI/P)

Nominal Concentration	Days
	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
Control	0.0	0.0	11.4	11.5	11.5	34.9	38.5	38.5	58.5	68.8	68.8	73.6	90.5	90.5	90.5	107.5
0.150 mg/L	0.0	0.0	9.3	10.9	10.9	31.4	38.1	38.1	55.6	68.0	68.0	74.0	90.3	90.3	90.3	102.8
0.470 mg/L	0.0	0.0	11.2	12.3	14.0	39.7	46.8	46.8	62.8	75.5	75.5	80.4	100.4	100.4	100.4	116.9
1.50 mg/L	0.0	0.0	8.6	8.7	10.5	34.7	34.7	34.7	53.0	56.8	56.8	72.5	82.6	82.6	87.0	104.5
4.70 mg/L	0.0	0.0	6.5	10.8	12.6	33.8	36.9	40.6	54.4	70.3	74.6	80.6	97.8	97.8	103.8	118.8
15.0 mg/L	*	*	*	*	*	*	*	*	*	*	*	*	*	*	*	*
*	All parentalDaphniawere dead during the 21-day testing period

The median lethal concentration (LC50) for parental Daphnia after the 21 -day exposure period is shown in Table 5 (below):

Table 5 - Calculated LC50 value for Parental Daphnia

Exposure Period (Days)	LC50*^1(mg/L)	95% Confidence Limits (mg/L)	Statistical Method
21	6.77	1.50 – 15.0	Binomial
*1	Based on the nominal concentration

The 50% reproduction inhibition concentration (EC50) after 21 -day exposure is shown in Table 6 (below):

Table 6 - Calculated EC50 Values for Inhibition of Reproduction

Exposure Period (Days)	EC50*^1(mg/L)	95% Confidence Limits (mg/L)	Statistical Method
21	8.40	--	Logit
*1	Based on the nominal concentration
--	Could not be determined

The maximum No Observed Effect Concentration (NOEC) and the Lowest Observed Effect Concentration (LOEC) for cumulative number of juveniles produced per adult alive are shown in Table 7 (attached).

Temperature of the test solution, dissolved oxygen concentration, pH and hardness during the exposure period are shown in Tables 8, 9, 10 and 11 respectively (attached).

Validity criteria fulfilled:

yes

Conclusions:

The median lethal concentration of parent Daphnia (LC50) was determined to be 6.77 mg/L with a 95 % confidence limit of 1.50 ~ 15.0 mg/L.

The 50% reproduction inhibition concentration (EC50) was determined as 8.40 mg/L although the confidence limit could not be calculated.

The maximum No Observed Effect Concentration (NOEC) was calculated as 4.70 mg/L and the Lowest Observed Effect Concentration (LOEC) was calculated as 15.0 mg/L.

Description of key information

LC50 (21 d)  6.77 mg/L for Daphnia magna (OECD 211)

EC50 (21 d) 8.40 mg/L based on Daphnia magna reproduction (OECD 211)

LOEC (21 d) 15.0 mg/L for Daphnia magna (OECD 211)

NOEC (21 d) 4.70 mg/L for Daphnia magna (OECD 211)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

4.7 mg/L

Additional information

According to ECHA guidance on information requirements and chemical safety assessment Chapter R.10: Characterisation of dose [concentration]-response for environment, two long-term results (such as EC10 or NOECs) from species representing two trophic levels (fish and/or Daphnia and/or algae) permit an assessment factor of 50 to be applied when calculating the freshwater PNEC.

The availability of a GLP study with analytical monitoring, which describes long-term toxicity to Daphnia magna (Ministry of Environment, Japan, 2000), allows that report to be used in conjunction with the most reliable long-term toxicity data from algae (Ministry of Environment, Japan, 1996) during PNEC calculation.