(E)-3-(benzyl(4-((4-nitrophenyl)diazenyl)phenyl)amino)propanenitrile

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:

supporting study

Study period:

14 November 2001 to 05 December 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to recent OECD test guidance in compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
None

Analytical monitoring:

not specified

Details on sampling:

Sampling was not undertaken due to the low water solubility; all results are based on the initial solubility assessment of 1 mg/l nominal.

Vehicle:

no

Details on test solutions:

To produce the only test concentration, 1 mg of the test substance was added to 1 litre of dilution water, treated for 1 hour with an ultrasonic bath followed by 24 hours stirring with a magnetic stirrer. The resulting solution was then filtered to remove undissolved particulates.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Source: Strain of Bundesgsundheitsamt Berlin
- Feeding during test: Yes
- Food type: Desmodesmus subspicatus
- Amount: day 1-7: 0.1 mg C/daphnia/day, day 8-21: 0.2mg C/daphnia/day
- Frequency:preferably daily but at least 3 times per week

ACCLIMATION
- Acclimation conditions (same as test or not): Yes
- Type and amount of food: Desmodesmus subspicatus
- Feeding frequency: ad libitum
- Health during acclimation (any mortality observed): Moratilties of parent Daphnia during culture period recorded daily in a semi-quantative way

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Post exposure observation period:

No post exposure observation period

Hardness:

The hardness of the dilution water ranged from 14.4-15.3 dH (- 257 - 273.1 mg/1 CaCO3) during the test.

Test temperature:

The temperature ranged from 19.1 - 20.7 deg C during the test.

pH:

The pH ranged from 7.8 - 8 during the test.

Dissolved oxygen:

8.4 - 8.5 mg/l

Salinity:

Not applicable.

Nominal and measured concentrations:

1 mg/l (nominal)

Details on test conditions:

TEST SYSTEM
- Test vessel: Beaker
- Material, size, headspace, fill volume: Glass, 150ml
- Aeration: None
- Renewal rate of test solution (frequency/flow rate): semi-static (renewal of test media on Monday, Wednesday, Friday) respectively.
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

OTHER TEST CONDITIONS
- Photoperiod: 16h light/8h darkness
- Light intensity: not exceeding 800 lux

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

- Mortality of parent animals: 0
- No. of offspring produced per day per female: see table
- Number of males and females (parental): All parthonogenic females
- Time to first brood release or time to hatch: first brood day 6
- Brood size: see table

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

The reproductive output of the Daphnia exposed to the test substance was compared to that of the control in order to determine the Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) according to DUNNETT (1955, 1964).

Reproduction rate
Nominal concentration |rag/1]: 1

Day	Date 2001	Number of living offspring per replicate
		1	2	3	4	5	6	7	8	9	10
0	Nov. 14
1	Nov.15
2	Nov. 16
3	Nov. 17
4	Nov. 18
5	Nov.19
6	Nov. 20	#	#	#	#	#	#	#	#	#	#
7	Nov. 21
8	Nov. 22		13	16							14
9	Nov. 23	8			7	S	12		5
10	Nov. 24									13
11	Nov. 25	9	4	10	7	11	8	16			9
12	Nov. 26								11
13	Nov. 27	33	32				2S	27		25
14	Nov. 28			34	36	30					34
15	Nov. 29								25
16	Nov. 30							29		29
17	Dec. 01	25	35	2S	39	26	33		37		33
18	Dec. 02
19	Dec. 03									24
20	Da: 04	34	27	22		28	29	22			19
21	Dec. 05				24				18
		109	111	110	113	103	110	94	96	91	109

# - first brood recognizableinbrood pouch of parem Daphnia

Validity criteria fulfilled:

yes

Conclusions:

Based on diflerent biological effects observed and endpoints assessed, the following results were obtained at the end of the 21 day exposure period of the Daphnia reproduction test:

Results [mg/l]
21d-NOEC (reproduction) : ≥ 1
21il-LOEC (reproduction) : > 1
21d-NOEC (mortality): ≥ 1
21d-LOEC (mortality) : > 1
1st brood on day: 6

Considering the low aqueous solubility of the test substance (< 1 mg/L) under exposure conditions no specific analysis was established; the above values refer to nominal concentrations.

Executive summary:

A study was performed to assess die effects of T-9601 on the reproductive output of parthenogenetic Daphnia magna STRAUS under semi-static conditions. The study was conducted in accordance with the OCCD Test guideline 211(Daphnia magna Reproduction Test; adopted: Sept. 21, 1998).

Young female Daphnia, aged less than 24 hours at the start of the test, were exposed to a limit test concentration of nominally 1 mg/L of T-9601 dissolved in water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and folded filters.

After a 21 d-exposure period, the total number of living offspring produced per parent animal alive at the end of the test was assessed.The reproductive output of the Daphnia exposed to the test substance was compared to that of the control in order to determine the Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) according to DUNNETT (1955, 1964). Additionally, the survival of the parent Daphnia and the time to production of first brood was determined.

 

Results [mg/L]

 

21d-NOEC (reproduction) : ≥ 1

21d-LOEC (reproduction): > 1

21d-NOEC(mortalility): ≥ 1

21d-LOEC(mortalility): > 1

1^st brood on day:     6

 

Considering the low aqueous solubility of the test substance (< 1 mg/L) under exposure conditions no specific analysis was established; the above values refer to nominal concentrations.

The test substance is not considered to be harmful to Daphnia via prolonged exposure at the limit of solubility in water.

Description of key information

Based on diflerent biological effects observed and endpoints assessed, the following results were obtained at the end of the 21 day exposure period of the Daphnia reproduction test:

Results [mg/L]
21d-NOEC (reproduction) : ≥ 1
21d-LOEC (reproduction) : > 1
21d-NOEC (mortality) : ≥ 1
21d-LOEC (mortality) : > 1
1st brood on day : 6

Considering the low aqueous solubility of the test substance (< 1 mg/L) under exposure conditions no specific analysis was established; the above values refer to nominal concentrations.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Remarks:

mortality and reproduction

Effect concentration:

>= 1 mg/L

Additional information

A study was performed to assess die effects of T-9601 on the reproductive output of parthenogenetic Daphnia magna STRAUS under semi-static conditions. The study was conducted in accordance with the OCCD Test guideline 211(Daphnia magna Reproduction Test; adopted: Sept. 21, 1998).

Young female Daphnia, aged less than 24 hours at the start of the test, were exposed to a limit test concentration of nominally 1 mg/L of T-9601 dissolved in water. Auxiliaries used to prepare the test media were an ultrasonic bath, a magnetic stirrer and folded filters.

After a 21 d-exposure period, the total number of living offspring produced per parent animal alive at the end of the test was assessed.The reproductive output of the Daphnia exposed to the test substance was compared to that of the control in order to determine the Lowest Observed Effect Concentration (LOEC) and the No Observed Effect Concentration (NOEC) according to DUNNETT (1955, 1964). Additionally, the survival of the parent Daphnia and the time to production of first brood was determined.

 

Results [mg/L]

 

21d-NOEC (reproduction) : ≥ 1

21d-LOEC (reproduction): > 1

21d-NOEC (mortalility): ≥ 1

21d-LOEC (mortalility): > 1

1^st brood on day:     6

 

Considering the low aqueous solubility of the test substance (< 1 mg/1) under exposure conditions no specific analysis was established; the above values refer to nominal concentrations.

The test substance is not considered to be harmful to Daphnia via prolonged exposure at the limit of solubility in water.