4,4'-bis(dimethylamino)-4''-(methylamino)trityl alcohol

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Justification for type of information:

Experimental test result performed according to the OECD test guideline.

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

- Concentrations: 0, 0.13, 0.2, 0.3, 0.45, 0.68 mg/l
- Sampling method: Samples for analytical determination was collected from four corners and center of the aquarium for each test concentrations send for chemical analysis
- Sample storage conditions before analysis: Samples were immediately analysed after sampling

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test chemical was prepared by dissolving 5000 mg of test chemical in 5000 ml of RO water. After stirring the stock solution was filtered and analytically detected and the concentration found to be 1000 mg/L. The final solubility value obtained after analytical detection was 10.55 mg/l. The remaining test solutions were prepared by dilution from the above stock solution. Test chemical concentrations used for the study were 0.13, 0.2, 0.3, 0.45, 0.68 mg/l, respectively.
- Controls: RO water as control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): Not applicable
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): Not applicable

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra Fish
- Length at study initiation (length definition, mean, range and SD): 2.3 cm
- Housing:The fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration.
- Duration of housing: 12 days
- Photoperiod: 16 hour light and 8 hour darkness

ACCLIMATION
- Acclimation period: Fishes, to be used in this study, are kept in the test water for seven days immediately before testing
- Acclimation conditions (same as test or not): same test conditions
Photoperiod: 16 hour light and 8 hours dark
Temperature range 21.4°C
pH range 8.0
DO range 7.2 mg/L
Feeding Daily once but not given atleast 48 hours before study
Aeration Continuous
Mortality (%): 0%
- Diet : Standard brand feed, once daily

FEEDING DURING TEST
- Food type: Feeding was not provided during test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test temperature:

21.4°C

pH:

8.0

Dissolved oxygen:

7.2 mg/l

Nominal and measured concentrations:

Test chemical concentrations used for the study were 0, 0.13, 0.2, 0.3, 0.45, 0.68 mg/l (Factor of 1.5), respectively.

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquarium
- Material, size, headspace, fill volume: 7 liters of test vessel filled with 4 liter of water
- Aeration: Aeration in test vessels was provided 1 day before the start of experiment
- No. of organisms per vessel: 7 test organisms per vessel
- No. of vessels per concentration (replicates): one vessel per concentrations
- No. of vessels per control (replicates): 1 vessel per control
- No. of vessels per vehicle control (replicates): Not applicable
- Biomass loading rate: 7 fishes /4 L

OTHER TEST CONDITIONS
- Photoperiod: 16 hours light and 8-hour darkness

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: RO water obatined from the RO water filter systems

OTHER TEST CONDITIONS
- Adjustment of pH: Not adjusted
- Photoperiod: 16 hours light and 8 hours dark

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Not applicable

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.257 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

No toxic effect was observed against fish at the limit test mg/L of water solubility under exposure conditions.
After the 96 hours of exposure, there was no mortality in the control group, and the dissolved oxygen concentration was 85.06% of the air saturation value.

Results with reference substance (positive control):

Not applicable

Sublethal observations / clinical signs:

OBSERVATIONS

Table: Assessment of test concentrations

 

Sr. No	Concentrations mg/L	Wavelength (nm)	Absorbance	Temperature (°C)
1	0.00	584.0	0.00	25
2	0.10	584.0	0.02	25
3	0.25	584.0	0.04	25
4	0.50	584.0	0.07	25
5	1.00	584.0	0.15	25
6	1.25	584.0	0.19	25
7	1.50	584.0	0.24	25
8	1.75	584.0	0.26	25
9	2.00	584.0	0.30	25
10	2.25	584.0	0.34	25
11	2.50	584.0	0.39	25
12	0.75	584.0	0.12	25

Table: Measured Concentration after analytical Determination

Sr. No	Nominal Conc.	0 Hr		96Hr
		Absorbance(mean)	Measured conc.	Absorbance(mean)	Measured conc.
1	Blank	0.00	0.00	0.00	0.00
2	0.13	0.002	0.01	0.008	0.05
3	0.2	0.03	0.19	0.03	0.22
4	0.3	0.04	0.25	0.03	0.23
5	0.45	0.05	0.34	0.04	0.23
6	0.68	0.08	0.50	0.07	0.46

The test concentrations were measured and found to remain 120-80% of nominal. Hence, the LC₅₀was expressed based on Nominal concentrations by probit analysis.

Mortality and symptoms

Sr. No	Test concentrations(mg/L)	3 hr	6 hr	24 hr	48 hr	72 hr	96 hr	Cumulative Mortality	Percent Inhibition
1	control	-	-	-	-	-	-	-	-
2	0.13	-	-	-	-	-	-	-	-
3	0.2	-	-	-	-	2	1	3	42.9
4	0.3	-	-	-	-	3	1	4	57.1
5	0.45	-	-	-	3	2	1	6	85.7
6	0.68	-	-	-	3	4	-	7	100

Visible Symptoms: 

Sr. No	Test concentrations(mg/L)	3 hr	6 hr	24 hr	48 hr	72 hr	96 hr
1	control	NS7	NS7	NS7	NS7	NS7	NS7
2	0.13	NS7	NS7	M7	M7	M7	M7
3	0.2	NS7	NS7	M7	M7	M5	M4
4	0.3	NS7	NS7	M7	M7	M4	M3
5	0.45	NS7	NS7	M7	M4 SC4	M2	M1
6	0.68	NS7	NS7	M7	M4 SC4	-	-
Any other symptoms		-		Purple stain is observed on fish body because of CAS	Purple stain is observed on fish body because of CAS	Purple stain is observed on fish body because of CAS	Purple stain is observed on fish body because of CAS

Explanation of symptoms:

A:apathy

N:Narcotic-like state

C:Swimming near water surface

M: Slow movements compared to control

NS:No abnormal symptoms 

S:Surfacing behaviour

L:Loss of equilibrium

SM:Shimming movement

H1:Hyperactivity

H2:Hypo activity

B:Bending behavior

F:Fin rot

IS:Invert Swimming

SB:Swimming behaviour

RF:Respiratory Function

P:Pigmentation

 

pH and oxygen content:

Test Concentration(mg/L)	pH
	24 Hours	48 Hours	72 Hours	96 Hours
control	7.9	7.1	7.9	8.0
0.13	8.1	7.9	8.1	8.1
0.2	8.4	8.0	8.2	8.2
0.3	8.3	8.1	8.2	8.2
0.45	8.3	8.1	8.2	8.1
0.68	8.3	8.1	-	-

 

Test Concentration(mg/L)	Dissolved Oxygen
	24 Hours	48 Hours	72 Hours	96 Hours
control	7.6	6.8	7.5	7.6
0.13	7.6	6.4	7.0	6.9
0.2	7.7	6.9	7.1	6.6
0.3	7.0	6.9	7.2	6.8
0.45	7.9	7.4	7.4	6.2
0.68	7.8	7.6	-	-

 

Test temperature

Test Concentration(mg/L)	Temperature °C
	24 Hours	48 Hours	72 Hours	96 Hours
control	21.3	21.1	21.1	22.6
0.13	21.3	21.1	21.1	22.6
0.2	21.3	21.1	21.1	22.6
0.3	21.3	21.1	21.1	22.6
0.45	21.3	21.1	21.1	22.6
0.68	21.3	21.1	-	-

Validity criteria fulfilled:

yes

Remarks:

*there was no mortality in the control group, and * the dissolved oxygen concentration was 85.06% (i.e. above 60%) of the air saturation value throughout the exposure period, test chemical conc. were analytically verfiied.

Conclusions:

Based on measured concentrations, the median lethal concentration [LC50 (96 h)] for test chemical on Danio rerio (Zebra Fish) was determined to be 0.257 mg/L (nominal conc.).

Executive summary:

An acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical on Zebra fish (Danio rerio). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average length of 2.3 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. Aeration in test vessels was provided till 1 day before the start of the experiment. The test chemical was prepared by dissolving 5000 mg of test chemical in 5000 ml of RO water. After stirring the stock solution was filtered and analytically detected and the concentration found to be 1000 mg/L.  The remaining test solutions were prepared by dilution from the above stock solution. Test chemical concentrations were analytically determined by UV-VIS spectrophotometer. Test chemical conc. used for the study were 0, 0.13, 0.2, 0.3, 0.45, 0.68 mg/l (factor of 1.5, respectively). Total 7 fishes were exposed to test chemical in a 7 lit of glass aquarium containing 4000 ml of potable water. The test vessels were placed in a room at a temperature of 21.4°C, pH of 8.0 and DO of 7.2 mg/l and under a photoperiod of 16:8 hr light: dark conditions, respectively. At regular intervals of 24 hours vessels were observed for any behavioural changes and mortality along with that the DO, pH and temperature was measured. Test chemical was sampled from test vessel for analytical determinations at 0 hour and 96 hours. All the test concentrations were analytical determined and at 96 hours of the exposure durations which were maintained in the range of 80 -120%. Therefore, the analysis of the results was based on nominal concentration. After the 96 hours of exposure, there was no mortality in the control group, and the dissolved oxygen concentration was 85.06% (i.e. above 60%) of the air saturation value throughout the exposure period. Thus, the study fulfilling the validity criterion as mentioned in the OECD TG 203. On the basis of effect of test chemical on mortality of the test organism, the 96 hr median lethal concentration [LC50 (96 h)] for test chemical on Danio rerio (Zebra Fish) was determined to be 0.257 mg/L (nomnal conc.). Thus, based on LC50 value, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic acute/ chronic 1' category as per the CLP classification criteria.

Description of key information

An acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical on Zebra fish (Danio rerio). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average length of 2.3 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. Aeration in test vessels was provided till 1 day before the start of the experiment. The test chemical was prepared by dissolving 5000 mg of test chemical in 5000 ml of RO water. After stirring the stock solution was filtered and analytically detected and the concentration found to be 1000 mg/L.  The remaining test solutions were prepared by dilution from the above stock solution. Test chemical concentrations were analytically determined by UV-VIS spectrophotometer. Test chemical conc. used for the study were 0, 0.13, 0.2, 0.3, 0.45, 0.68 mg/l (factor of 1.5, respectively). Total 7 fishes were exposed to test chemical in a 7 lit of glass aquarium containing 4000 ml of potable water. The test vessels were placed in a room at a temperature of 21.4°C, pH of 8.0 and DO of 7.2 mg/l and under a photoperiod of 16:8 hr light: dark conditions, respectively. At regular intervals of 24 hours vessels were observed for any behavioural changes and mortality along with that the DO, pH and temperature was measured. Test chemical was sampled from test vessel for analytical determinations at 0 hour and 96 hours. All the test concentrations were analytical determined and at 96 hours of the exposure durations which were maintained in the range of 80 -120%. Therefore, the analysis of the results was based on nominal concentration. After the 96 hours of exposure, there was no mortality in the control group, and the dissolved oxygen concentration was 85.06% (i.e. above 60%) of the air saturation value throughout the exposure period. Thus, the study fulfilling the validity criterion as mentioned in the OECD TG 203. On the basis of effect of test chemical on mortality of the test organism, the 96 hr median lethal concentration [LC50 (96 h)] for test chemical on Danio rerio (Zebra Fish) was determined to be 0.257 mg/L (nomnal conc.). Thus, based on LC50 value, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic acute/ chronic 1' category as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.257 mg/L

Additional information

An acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical on Zebra fish (Danio rerio). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Zebra fish (Danio rerio) of average length of 2.3 cm was used as a test organism for the study. Test fishes were kept in a static tank in tap water passed through reverse osmosis system, under natural conditions along with proper feed and aeration. During the housing period, test fishes were fed once daily with standard brand fed. Aeration in test vessels was provided till 1 day before the start of the experiment. The test chemical was prepared by dissolving 5000 mg of test chemical in 5000 ml of RO water. After stirring the stock solution was filtered and analytically detected and the concentration found to be 1000 mg/L.  The remaining test solutions were prepared by dilution from the above stock solution. Test chemical concentrations were analytically determined by UV-VIS spectrophotometer. Test chemical conc. used for the study were 0, 0.13, 0.2, 0.3, 0.45, 0.68 mg/l (factor of 1.5, respectively). Total 7 fishes were exposed to test chemical in a 7 lit of glass aquarium containing 4000 ml of potable water. The test vessels were placed in a room at a temperature of 21.4°C, pH of 8.0 and DO of 7.2 mg/l and under a photoperiod of 16:8 hr light: dark conditions, respectively. At regular intervals of 24 hours vessels were observed for any behavioural changes and mortality along with that the DO, pH and temperature was measured. Test chemical was sampled from test vessel for analytical determinations at 0 hour and 96 hours. All the test concentrations were analytical determined and at 96 hours of the exposure durations which were maintained in the range of 80 -120%. Therefore, the analysis of the results was based on nominal concentration. After the 96 hours of exposure, there was no mortality in the control group, and the dissolved oxygen concentration was 85.06% (i.e. above 60%) of the air saturation value throughout the exposure period. Thus, the study fulfilling the validity criterion as mentioned in the OECD TG 203. On the basis of effect of test chemical on mortality of the test organism, the 96 hr median lethal concentration [LC50 (96 h)] for test chemical on Danio rerio (Zebra Fish) was determined to be 0.257 mg/L (nomnal conc.). Thus, based on LC50 value, test chemical was considered as toxic to aquatic fishes and hence, considered to be classified in 'aquatic acute/ chronic 1' category as per the CLP classification criteria.