Tetrasodium 4,4'-bis[[4-morpholino-6-(p-sulphonatoanilino)-1,3,5-triazin-2-yl]amino]stilbene-2,2'-disulphonate

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:

Experiment start date -13 May 1992; Experiment end date-03 June 1992; Study completion date - 14 September 1992.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Read-across based on grouping of substances (category approach). A detailed category justification is included in chapter 13 of this dossier.

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: internal company procedure.
- Derived solution: 1500 mg/l

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea, parthenogenetic females.
- Colony: clone strain from Bundesgesundheitsamt Berlin.
- Source: Bayer AG Leverkusen Laboratories.
- Age: 6 - 24 hours.
- Feeding during test: 0.2 mg/adult daphnia (day 0-3); 0.4 mg/adult daphnia (day 4-7); 0.6 mg/adult daphnia (day 8-21)
- Food type: Scenedesmus subspicatus from fermenter culture. The concentrated algal suspension is prepared fresh weekly and stored maximum of 7 days in the refrigerator. When the algae tend, when fed to the formation of precipitates, a 5-minute ultrasound treatment is recommended.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Test temperature:

20 ± 2 °C (maximum in individual test: ± 1 °C).

pH:

7.6 - 8.8

Dissolved oxygen:

87.3 - 128.5 %

Nominal and measured concentrations:

10, 31.6, 100, 316 and 1000 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: beakers (diameter 5.5 cm, high 9.5 cm), with a coverage of 6 cm of diameter.
- Fill volume: 100 ml/beaker.
- Aeration: not aerated.
- Renewal rate of test solution: thrice a week (on Monday, Wednesday and Friday).
- No. of organisms per concentration: 10 (both tested and control groups).
- Replicates: 10 per concentration.

TEST MEDIUM
- Intervals of water quality measurement: once a week.

OTHER TEST CONDITIONS
- Photoperiod: 16 h light / 8 h darkness
- Light intensity: ca. 1700 lux
- Irradiation: 25 µE/m^2 x s

EFFECT PARAMETERS MEASURED: reproduction, mobility.

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

6.59 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

31.6 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Immoobilization EC50 (21d): 31.6 -100 mg/l (nominal)
Reproductive EC50 (21d): 30.2 mg/l (probit-analysis based on nominal concentrations).

TEST RESULTS

Conc (mg/l)	Immobilization/mortality of parents at 21 day		Reproduction rate (R21)
			Pup/parent	%
Control	0	0	92.0	100
10	0	0	91.6	99.6
31.6	0	0	42.1	45.8
100	10	100	0.0	0.0
316	10	100	0.0	0.0
1000	10	100	0.0	0.0

VARIABILITY of REPRODUCTION FROM CONTROL TO TEST CONCENTRATION

Concentration (mg/l)	Reproduction rate at the 21th day	Standard Deviation	Variation coefficient (%)
Control	92.0	11.9	12.9
10	91.6	10.2	11.1
31.6	42.1	22.0	52.3
100	0.0	0.0	0.0
316	0.0	0.0	0.0
1000	0.0	0.0	0.0

 

Control

Day	Puppies										Stillbirths
	1	2	3	4	5	6	7	8	9	10	3	5	6	7
0
1
2
3
4
5
6			#	#			#	#
7	#	#			#	#			#	#
8
9	10							15	10	15
10
11
12	5	16	15	10	11	16	17	15	18	13
13
14	15	22	17	16		25	16	10	1	2	2T	1T	1T	1T
15
16	16	30	18	10	33	7	11	8	30	30
17
18
19	20	20	18	21	13	22	16	11	33	8
20
21	26	21	26	23	15	32	24	28		40
Reproduction rate at the 21th day	92	109	94	80	72	102	84	87	92	108	2T	1T	1T	1T

#: First pups recognizable in the mother's brood chamber.

T: dead pup

Concentration of 10 mg/l

Day	Puppies										Stillbirths
	1	2	3	4	5	6	7	8	9	10	4	8	9	10
0
1
2
3
4
5
6		#	#	#	#	#	#	#	#	#
7	#
8
9	13	20	14	20	22		18	23	15	12
10
11
12	14	23	8			18		16	20	10				10T
13
14	4	2	20	23	21	23	20				1T
15
16	25	28					8	33	26	20				7T
17
18
19	31		23	20	21	15	16	28	24	30		1T	3T
20
21	28	15	33	24	26	26	27			10
Reproduction rate at the 21th day	115	88	98	87	90	82	89	100	85	92	1T	1T	3T	17T

#: First pups recognizable in the mother's brood chamber.

T: dead pup

Concentration of 31.6 mg/l

Day	Puppies
	1	2	3	4	5	6	7	8	9	10
0
1
2
3
4
5
6			#	#	#		#	#
7	#
8
9		#				#			#	#
10
11
12	6	6	15	18	5	11	10	10	2
13
14	23		28	31					5
15
16	6	10			10	20	10	8		11
17
18
19	8	16	5	16	4	22			7	15
20
21	25		16	15	6		6	10	5
Reproduction rate at the 21th day	68	32	64	80	25	53	26	28	19	26

#: First pups recognizable in the mother's brood chamber.

T: dead pup

Concentration of 100 mg/l

Day	Puppies
	1	2	3	4	5	6	7	8	9	10
0
1
2
3
4
5				+	+	+	+	+	+	+
6
7			+
8
9	+	+
10
11
12
13
14
15
16
17
18
19
20
21
Reproduction rate at the 21th day

+: dead in the mother's brood chamber

Concentration of 316 mg/l

Day	Puppies
	1	2	3	4	5	6	7	8	9	10
0
1
2
3
4
5	+	+	+	+	+	+	+	+	+	+
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
Reproduction rate at the 21th day

+: dead in the mother's brood chamber

Concentration of 1000 mg/l

Day	Puppies
	1	2	3	4	5	6	7	8	9	10
0
1
2
3
4
5	+	+	+	+	+	+	+	+	+	+
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
Reproduction rate at the 21th day

+: dead in the mother's brood chamber

MEASURED CONCENTRATIONS

Sampling time	Concentration
	Nominal	Measured (mg/l)	%
Initial condition	Control	< 0.2	-
	10	6.4	64
	100	804.7	80.5
48 hrs	Control	< 0.2	-
	10	7.2	72
	100	632.6	63.3
Renewal	Control	< 0.2	-
	10	6.5	65
	100	727.4	72.7
72 hrs	Control	< 0.2	-
	10	6.3	63
	100	667.7	66.8

Validity criteria fulfilled:

yes

Remarks:

The mortality of the parent animals does not exceed 20 % at the end of the test, in the control; nevertheless there are no information about the mean number of live offspring produced per parent animal surviving at the end of the test

Conclusions:

The reproductive NOEC (21 d) was found to be 6.59 mg/l (geom. mean).

Executive summary:

Method

The test substance chronic toxicity on aquatic invertebrates (Daphnia Magna) was evaluated according to the OECD guideline 211.

The nominal concentrations used were: 10, 31.6, 100, 316 and 1000 mg/l.

Results

Reproductive NOEC (21 d): 6.59 mg/l (geom. mean)

Mortality/immobilization NOEC (21 d): 31.6 mg/l (nominal)

Description of key information

NOEC 21d daphnids = 6.59 mg/l

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

6.59 mg/L

Additional information

A Daphnia magna reproductive study is available on the analogous substance CAS 16470-24-9 with NOEC reported as geometric mean because the difference between nominal and measured values is often greater than ± 20 %. The test was performed on a similar substance within the category of Stilbene Fluorescent Whitening Agents, the analogous dihydroxyethyl derivative tetrasulphonated sodium salt. The diethylamino substituent is less polar, less reactive and makes the molecule slightly less soluble compared to the dihydroxyethylamino derivative. The latter can therefore considered as a representative conservative analogous for the aquatic toxicity because of the higher bioavailability and higher reactivity with organic substrates. The Category Justification Report is attached to the section 13.