N-[4-[(9,10-dihydro-4-hydroxy-9,10-dioxo-1-anthryl)amino]phenyl]acetamide

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:

guideline study

Justification for type of information:

Data is from experimental study report

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

- Pre-treatment of test chemical: The in-house water solubility for the test chemical is 0.22 mg/L. Therefore, the test chemical will be prepared by dissolving 1000 mg of test chemical in 1000 mL of M7 media with 48 hours stirring to get the final concentration of 1000 mg/L. This stock solution will be filtered by using whatman filter paper no. 42, which will then analytically determine. The final solubility value obtained in media was used to prepare the remaining test concentrations from the above stock solution.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Test chemical was weighed and about 1000mg in 1000 ml of test chemical kept for stirring for 48 hours, which was then filtered and the concentration was anlytically determined, and which was obtained to be 0.1 mg/L
- Controls: Test media M7 was directly used as the control.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): no vehilce was used during study
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): no applicable

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Age at study initiation (mean and range, SD): < 24 hr old
- Length at study initiation (length definition, mean, range and SD): 0.45 cm
- Source: Eggs of Daphnia magna were obtained from Denmark Technical University and maintained at Unique Ecotox Research Laboratory, Nagpur

ACCLIMATION
- Type and amount of food: Living cells of Selenestrum capricornutum
- Feeding frequency: Feeding of the Daphnia with living cells of Selenestrum capricornutum was done thrice a week.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Hardness:

140 mg/l as CaCO3

Test temperature:

18-22°C

pH:

7.4-7.9

Dissolved oxygen:

7.4-8.1mg/L

Salinity:

Not applicable

Nominal and measured concentrations:

0.1 mg/l ( nominal concentration)

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker
- Material, size, headspace, fill volume: 50 ml
- Aeration: Test vessel were not aerated during the study.
- No. of vessels per concentration (replicates):
- No. of vessels per control (replicates):
- No. of vessels per vehicle control (replicates):

TEST MEDIUM / WATER PARAMETERS: M7 medium

OTHER TEST CONDITIONS
- Photoperiod: 16:8 light:dark conditions
- Light intensity: Not exceeding 1000-1500 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia.

Reference substance (positive control):

not specified

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

0.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

- Behavioural abnormalities: no behavioural abnormalities were observed
- Observations on body length and weight: no significant difference was observed in the body length was observed in both parents and offsprings
- Other biological observations: first brood with visible eggs in parents was observed on day 11
- Mortality of control: no moratality was observed in control parents and offsprings
- Other adverse effects control: no adverse effects were observed
- Immobilisation of control: no immobilisation

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The statistical comparision was done by student T test method using Sys stat software

Defined concentrations of the test chemical led to a certain percentage reduction of the parthenogenetic reproduction rate at the end of the 21 day study period. The living offspring was counted daily along with the renewal of the test medium. However, test vessels were inspected daily for the occurrence of juveniles and marked accordingly.

Control

Day	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
Medium renewal	-	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
pH (new)	7.4	-	7.9	-	7.3	-	7.8	-	7.8	-	-	7.4	-	7.3	-	7.5	-	-	7.4	-	7.5	-
O2 (mg/l)      (new)	7.5	-	8.0	-	7.8	-	7.9	-	7.8	-	-	7.8	-	7.9	-	7.6	-	-	7.9	-	7.7	-
Temp (°C) (new)	18	-	21	-	20.7	-	21.0	-	21.6	-	-	21.8	-	21.3	-	21.5	-	-	21.9	-	21.5	-
Food provided	Y	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
Replicates	Number of living offspring per replicate																						Total no. of live offsprings per vessel
V1	-	-	-	-	-	-	-	-	-	-	-	-	-	15	-	-	15	-	11	16	-	-	57
V2	-	-	-	-	-	-	-	-	-	-	-	-	10	-	-	26	-	-	9	-	-	-	45
V3	-	-	-	-	-	-	-	-	-	-	-	-	-	-	10	-	-	-	10	-	11	26	57
V4	-	-	-	-	-	-	-	-	-	-	-	#25	-	15	16	4	-	-	7	-	13	-	80
V5	-	-	-	-	-	-	-	-	-	-	-	7	-	20	-	-	21	-	12	7	-	-	67
V6	-	-	-	-	-	-	-	-	-	-	-	15	-	25	-	-	-	-	16	-	14	-	70
V7	-	-	-	-	-	-	-	-	-	-	-	-	15	-	6	-	19	-	21	-	21	-	82
V8	-	-	-	-	-	-	-	-	-	-	-	-	-	11	-	-	15	-	12	15	-	-	53
V9	-	-	-	-	-	-	-	-	-	-	-	-	18	-	-	-	13	-	7	11	-	-	49
V10	-	-	-	-	-	-	-	-	-	-	-	-	10	-	5	-	-	-	21	-	-	16	52
Cumulative Reproductivity																							612

No. of Days		0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
V1	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V2	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V3	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V4	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V5	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V6	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V7	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V8	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V9	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V10	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

Nominal Concentration: 0.10 mg/L

Day	0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
Medium renewal	-	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
pH (new)	7.5	-	7.9	-	7.5	-	7.9	-	7.8	-	-	7.5	-	7.4	-	7.5	-	-	7.4	-	7.8	-
O2 (mg/l)      (new)	8.1	-	8.0	-	7.9	-	8.0	-	7.4	-	-	7.9	-	7.9	-	8.0	-	-	7.7	-	7.4	-
Temp (°C) (new)	18	-	21	-	21.2	-	21.0	-	21.8	-	-	21.8	-	21.0	-	21.5	-	-	21.9	-	21.7	-
Food provided	Y	-	Y	-	Y	-	Y	-	Y	-	-	Y	-	Y	-	Y	-	-	Y	-	Y	-
Replicates	Number of living offspring per replicate																						Total no. of live offspring’s per vessel
V1	-	-	-	-	-	-	-	-	-	-	-	#5	-	1	10	-	-	12	10	2	15	-	55
V2	-	-	-	-	-	-	-	-	-	-	-	-	5	1	12	5	-	-	12	5	5	-	45
V3	-	-	-	-	-	-	-	-	-	-	-	11	-	-	8	-	9	-	15	-	-	-	43
V4	-	-	-	-	-	-	-	-	-	-	-	1	-	-	6	12	-	-	5	-	14	16	54
V5	-	-	-	-	-	-	-	-	-	-	-	4	-	13	-	2	-	-	2	17	-	10	48
V6	-	-	-	-	-	-	-	-	-	-	-	10	-	-	25	-	-	-	7	-	13	12	67
V7	-	-	-	-	-	-	-	-	-	-	-	-	-	10	-	15	-	-	-	21	-	-	46
V8	-	-	-	-	-	-	-	-	-	-	-	-	-	8	-	2	-	-	9	25	2	15	61
V9	-	-	-	-	-	-	-	-	-	-	-	21	10	-	-	4	-	-	-	14	-	-	49
V10	-	-	-	-	-	-	-	-	-	-	-	-	-	3	-	14	-	-	-	10	-	21	48
Cumulative Reproductivity																							516

 

 

 

No. of Days		0	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21
V1	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V2	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V3	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V4	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V5	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V6	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V7	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V8	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V9	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
V10	Dead P	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
	Dead O	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-

 

Validity criteria fulfilled:

yes

Conclusions:

Based on the effect on reproduction of the test daphnids, the 21 d NOEC value was determined to be 0.1 mg/l.

Executive summary:

A chronic study was conducted for assesing the effect of test chemical. The study was performed in accordance with the principles of the OECD Guideline 211 (Daphnia magna Reproduction Test). Daphnia magna of length 0.45 cm was used as a test organism. Eggs of Daphnia magna were obtained from Denmark Technical University. Test daphnids were fed with living cells of Selenestrum capricornutum and it was done thrice a week. The in-house water solubility for the test chemical is 0.22 mg/L. Therefore, the test chemical will be prepared by dissolving 1000 mg of test chemical in 1000 mL of M7 media with 48 hours stirring to get the final concentration of 1000 mg/L. This stock solution will be filtered by using whatman filter paper no. 42, which will then analytically determine. The final solubility value obtained in media was used to prepare the remaining test concentrations from the above stock solution. The analytical determinations were performed by UV-VIS spectrophotometer. The pre-treated stock solution was then diluted with media in order to get the required test solutions ranging from concentrations, 0.1, 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, and 7.0 mg/L. The absorbance of resulting solution was measured using UV-VIS spectrophotometer against corresponding blank at lambda max (λmax). Standard curve was plotted against concentration verses absorbance and the maximum solubility was determined from the below standard curve. Analytical assessments were performed for selected test concentration at 1st, 2nd and 3rd week. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. Test chemical conc. used in the defiinite study was 0.1 mg/l. Thus, limit test was performed at 0.1 mg/l conc. Test daphnids were exposed to test chemical conc. in a glass beaker for an exposure period of 21 days. Vessels were not aerated during the study. M7 medium was used as a test medium. Test conditions involve a temperature range of 18-22°C, 16:8 light:dark conditions and light intensity not exceeding 1000-1500 lux, respectively. The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia. Defined concentrations of the test chemical led to a certain percentage reduction of the parthenogenetic reproduction rate at the end of the 21 day study period. The living offspring was counted daily along with the renewal of the test medium. However, test vessels were inspected daily for the occurrence of juveniles and marked accordingly. On the basis of the effect on reproduction of the test daphnids, the 21 d NOEC value was determined to be 0.1 mg/l. Thus based on the outcomes chemical could be classified into aquatic chronic category 1 as per CLP classification criteria.

Description of key information

Long term toxicity to aquatic invertebrate:

A chronic study was conducted for assesing the effect of test chemical. The study was performed in accordance with the principles of the OECD Guideline 211 (Daphnia magna Reproduction Test). Daphnia magna of length 0.45 cm was used as a test organism. Eggs of Daphnia magna were obtained from Denmark Technical University. Test daphnids were fed with living cells of Selenestrum capricornutum and it was done thrice a week. DMSO or acetone was used as a vehicle. The in-house water solubility for the test chemical is 0.22 mg/L. Therefore, the test chemical will be prepared by dissolving 1000 mg of test chemical in 1000 mL of M7 media with 48 hours stirring to get the final concentration of 1000 mg/L. This stock solution will be filtered by using whatman filter paper no. 42, which will then analytically determine. The final solubility value obtained in media was used to prepare the remaining test concentrations from the above stock solution. The analytical determinations were performed by UV-VIS spectrophotometer. The pre-treated stock solution was then diluted with media in order to get the required test solutions ranging from concentrations, 0.1, 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, and 7.0 mg/L. The absorbance of resulting solution was measured using UV-VIS spectrophotometer against corresponding blank at lambda max (λmax). Standard curve was plotted against concentration verses absorbance and the maximum solubility was determined from the below standard curve. Analytical assessments were performed for selected test concentration at 1st, 2nd and 3rd week. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. Test chemical conc. used in the defiinite study was 0.1 mg/l. Thus, limit test was performed at 0.1 mg/l conc. Test daphnids were exposed to test chemical conc. in a glass beaker for an exposure period of 21 days. Vessels were not aerated during the study. M7 medium was used as a test medium. Test conditions involve a temperature range of 18-22°C, 16:8 light:dark conditions and light intensity not exceeding 1000-1500 lux, respectively. The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia. Defined concentrations of the test chemical led to a certain percentage reduction of the parthenogenetic reproduction rate at the end of the 21 day study period. The living offspring was counted daily along with the renewal of the test medium. However, test vessels were inspected daily for the occurrence of juveniles and marked accordingly. On the basis of the effect on reproduction of the test daphnids, the 21 d NOEC value was determined to be 0.1 mg/l. Thus based on the outcomes chemical could be classified into aquatic chronic category 1 as per CLP classification criteria.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.1 mg/L

Additional information

Long term toxicity to aquatic invertebrate:

A chronic study was conducted for assesing the effect of test chemical. The study was performed in accordance with the principles of the OECD Guideline 211 (Daphnia magna Reproduction Test). Daphnia magna of length 0.45 cm was used as a test organism. Eggs of Daphnia magna were obtained from Denmark Technical University. Test daphnids were fed with living cells of Selenestrum capricornutum and it was done thrice a week. DMSO or acetone was used as a vehicle. The in-house water solubility for the test chemical is 0.22 mg/L. Therefore, the test chemical will be prepared by dissolving 1000 mg of test chemical in 1000 mL of M7 media with 48 hours stirring to get the final concentration of 1000 mg/L. This stock solution will be filtered by using whatman filter paper no. 42, which will then analytically determine. The final solubility value obtained in media was used to prepare the remaining test concentrations from the above stock solution. The analytical determinations were performed by UV-VIS spectrophotometer. The pre-treated stock solution was then diluted with media in order to get the required test solutions ranging from concentrations, 0.1, 0.25, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, and 7.0 mg/L. The absorbance of resulting solution was measured using UV-VIS spectrophotometer against corresponding blank at lambda max (λmax). Standard curve was plotted against concentration verses absorbance and the maximum solubility was determined from the below standard curve. Analytical assessments were performed for selected test concentration at 1st, 2nd and 3rd week. The concentration of the test chemical being tested has been satisfactorily maintained within ± 20 % of the nominal concentration throughout the test. Therefore, the analysis of the results was based on nominal concentration. Test chemical conc. used in the defiinite study was 0.1 mg/l. Thus, limit test was performed at 0.1 mg/l conc. Test daphnids were exposed to test chemical conc. in a glass beaker for an exposure period of 21 days. Vessels were not aerated during the study. M7 medium was used as a test medium. Test conditions involve a temperature range of 18-22°C, 16:8 light:dark conditions and light intensity not exceeding 1000-1500 lux, respectively. The evaluation of the NOEC was determined based on the number of offspring’s produced per living parent Daphnia. Defined concentrations of the test chemical led to a certain percentage reduction of the parthenogenetic reproduction rate at the end of the 21 day study period. The living offspring was counted daily along with the renewal of the test medium. However, test vessels were inspected daily for the occurrence of juveniles and marked accordingly. On the basis of the effect on reproduction of the test daphnids, the 21 d NOEC value was determined to be 0.1 mg/l. Thus based on the outcomes chemical could be classified into aquatic chronic category 1 as per CLP classification criteria.