Cinnamaldehyde

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:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

The method followed that described in the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Directive 92/69/EEC (which constitutes Annex V of Council Directive 67/548/EEC).

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

Council Directive 92/69/EEC C.1 (1992)

GLP compliance:

yes

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

Control : synthetic fresh water was used as media and prepared in accordance with ISO standards.
Water for dilution : Test chemical was directly weighed and dissolved in the synthetic fresh water prepared in accordance with ISO standards and spinned for 60 seconds at 8000 rpm with an ultra turrax.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species : Zebra fish (Brachydanio rerio)
Origin : west-aquarium
Body lenght : 2.5-3.5 cm
Mortality while kept : <2%
Keeping water : Synthetic fresh water in accordance with ISO.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

14.5 [°dH]

Test temperature:

20.7-22.8°C

pH:

7.5 - 8.2 pH

Dissolved oxygen:

8.2 - 9.6 (mg/l)

Nominal and measured concentrations:

Test concentration/s (mg/l) : 0, 2.8, 3.9, 5.5 and 7.8 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: test vessels (300 × 135 × 200 mm)
- Type (delete if not applicable): closed, yes test vessel was ventilated.
- Material, size, headspace, fill volume: 5 l
- Renewal rate of test solution (frequency/flow rate): Daily renewal of the test medium was done during the study.
- Other: Test chemical was solubilized using the Ultra-turrax

OTHER TEST CONDITIONS
Photoperiod : 12 hours light/12 hours dark

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

1.41 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.35 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 3.9 - < 5.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

5.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

3.93 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

2.8 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.3 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: plot of log conc vs % mortality

Reported statistics and error estimates:

The LC0 and the LC100 are determined directly from the study, while the LC50 is calculated or read from the dose-response curve.

Sublethal observations / clinical signs:

A defined concentration of the test substance will lead to a certain percentage proportion of dead fish at the end of the study period.The LC0 and the LC100 are determined directly from the study,while the LC50 is calculated or read from the dose-response curve.The study lasts for 96 hours.

Concentration: Control 

Abiotic parameter

	0 h 03-01	24 h 03-02	0 h 03-02	24 h 03-03	0 h 03-03	24 h 03-04	0 h 03-04	24 h 03-05
Temperature °C	22.6	21.8	21.3	21.8	20.7	21.6	20.8	21.6
Oxygen mg/L	8.9	8.4	8.6	8.9	9.1	8.4	8.9	8.6
Oxygen %  saturation	105.5	98.1	99.5	104.0	104.2	97.8	112.2	100.1
pH value	8.2	7.8	8.0	7.4	7.9	7.7	7.9	7.5

 

Mortality:

	2 h	24 h	48 h	72 h	96 h
Absolute	0	0	0	0	0
Cumulative	0	0	0	0	0
Cumulative (%)	0	0	0	0	0

 

Analysis:

Newly prepared media	93-03-01	93-03-02	93-03-03	93-03-04
Analytical values (mg/L)	Not detected <0.25	Not detected <0.25
Media after 24 hour exposure	93-03-02	93-03-03	93-03-04	93-03-05
Analytical values (mg/L)	Not detected <0.25	Not detected <0.25

Concentration: 2.8 mg/L

Abiotic parameters

	0 h 03-01	24 h 03-02	0 h 03-02	24 h 03-03	0 h 03-03	24 h 03-04	0 h 03-04	24 h 03-05
Temperature °C	22.7	21.7	21.4	21.8	20.9	21.7	21.0	21.9
Oxygen mg/L	9.1	8.2	9.1	9.2	9.5	9.0	9.1	8.7
Oxygen %  saturation	107.9	95.6	105.6	106.7	109.2	104.9	104.8	101.8
pH value	8.2	7.8	8.0	7.5	7.9	7.7	7.9	7.5

 

Mortality:

	2 h	24 h	48 h	72 h	96 h
Absolute	0	0	0	0	0
Cumulative	0	0	0	0	0
Cumulative (%)	0	0	0	0	0

 

Analysis:

Newly prepared media	93-03-01	93-03-02	93-03-03	93-03-04
Analytical values (mg/L)	2.68	2.73
Media after 24 hour exposure	93-03-02	93-03-03	93-03-04	93-03-05
Analytical values (mg/L)	0.87	1.94

 

Concentration: 3.9 mg/L

Abiotic parameters

	0 h 03-01	24 h 03-02	0 h 03-02	24 h 03-03	0 h 03-03	24 h 03-04	0 h 03-04	24 h 03-05
Temperature °C	22.7	21.8	21.3	21.8	21.4	21.8	21.1	21.9
Oxygen mg/L	9.6	8.4	9.3	9.2	9.3	8.9	9.2	8.7
Oxygen %  saturation	113.9	97.9	107.6	107.5	107.9	104.0	106.1	101.8
pH value	8.2	7.8	8.0	7.5	7.9	7.7	7.9	7.5

 

Mortality:

	2 h	24 h	48 h	72 h	96 h
Absolute	0	0	0	0	2
Cumulative	0	0	0	0	2
Cumulative (%)	0	0	0	0	20

 

Analysis:

Newly prepared media	93-03-01	93-03-02	93-03-03	93-03-04
Analytical values (mg/L)	3.75	3.69
Media after 24 hour exposure	93-03-02	93-03-03	93-03-04	93-03-05
Analytical values (mg/L)	2.12	2.62

 

  Concentration: 5.5 mg/L

Abiotic parameters

	0 h 03-01	24 h 03-02	0 h 03-02	24 h 03-03	0 h 03-03	24 h 03-04	0 h 03-04	24 h 03-05
Temperature °C	22.7	21.5	21.5	21.6	21.4	21.7	21.2	21.9
Oxygen mg/L	9.4	8.8	8.6	9.2	9.6	9.0	9.2	8.9
Oxygen %  saturation	111.5	102.2	99.9	107.1	111.4	104.9	106.4	104.1
pH value	8.2	7.7	8.0	7.6	8.0	7.7	7.9	7.8

 

Mortality:

	2 h	24 h	48 h	72 h	96 h
Absolute	0	0	0	9	1
Cumulative	0	0	0	9	10
Cumulative (%)	0	0	0	90	100

 

Analysis:

Newly prepared media	93-03-01	93-03-02	93-03-03	93-03-04
Analytical values (mg/L)	5.42	5.23
Media after 24 hour exposure	93-03-02	93-03-03	93-03-04	93-03-05
Analytical values (mg/L)	3.67	4.18

  Concentration: 7.8 mg/L

Abiotic parameters

	0 h 03-01	24 h 03-02	0 h 03-02	24 h 03-03	0 h 03-03	24 h 03-04	0 h 03-04	24 h 03-05
Temperature °C	22.8	21.7	21.5	21.1
Oxygen mg/L	9.5	8.5	8.6	9.0
Oxygen %  saturation	113.0	99.1	99.9	103.3
pH value	8.2	7.8	8.0	7.6

 

Mortality:

	2 h	24 h	48 h	72 h	96 h
Absolute	0	1	9
Cumulative	0	1	10
Cumulative (%)	0	10	100

 

Analysis:

Newly prepared media	93-03-01	93-03-02	93-03-03	93-03-04
Analytical values (mg/L)	7.43	7.09
Media after 24 hour exposure	93-03-02	93-03-03	93-03-04	93-03-05
Analytical values (mg/L)	5.54	6.72

 

 Note: analytical values determined over the first two 24 hours exposure periods are very similar, thus making it unnecessary to perform further determinations.

Validity criteria fulfilled:

yes

Remarks:

According to OECD 203 all validity criterias are met: -No mortality in control was observed - ASV> 60% - Analytical monitoring is also done.

Conclusions:

Based on the effect of test chemical on the mortality of the test organism, the 96hr LC0, LC50 and LC100 value was determined to be 1.41 mg/l (measured conc.), 2.35 mg/l (measured conc.) & 3.93 mg/l (measured conc.) and the 96 hr LC50 & LC100 value was determined to be in the range of > 3.9 to < 5.5 mg/l (nominal conc.) and 5.5 mg/l (nominal conc.), respectively.

Executive summary:

An acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical in fish. Study was performed following the EU Method C.1 (Acute Toxicity for Fish) (Council Directive 92/69/EEC C.1 (1992)). Danio rerio (Zebra fish) obtained from west-aquarium of 2.5 to 3.5 cm length was used as a test organism for the study. Test chemical concentrations were verified analytically by HPLC at the start and after 24 hr of the experiment. Nominal test chemical concentrations used for the study were 0, 2.8, 3.9, 5.5 and 7.8 mg/l, respectively. The test solution was preared by dissolved the test chemical into synthetic water and treated for 60 seconds at 8000 rpm with an ultra-turrax. Test fishes were exposed with the test chemical in a close ventilated test vessel. Study was performed using test fishes in a semi-static system at 20.7-22.8°C temperature, 7.5 - 8.2 pH, 8.2 - 9.6 (mg/l) dissolved oxygen and hardness 14.5 [°dH] under a photoperiod of 12:12 light:dark conditions for 96 hrs. Daily renewal of the test medium was done during the study. The LC0 and the LC100 are determined directly from the study, while the LC50 is calculated or read from the dose-response curve. On the basis of the effect of test chemical on the mortality of the test organism, the 24hr LC0, LC50 and LC100 value was determined to be 1.41 mg/l (measured conc.), 2.35 mg/l (measured conc.) & 3.93 mg/l (measured conc.) and the 96 hr LC50 & LC100 value was determined to be in the range of > 3.9 to < 5.5 mg/l (nominal conc.) and 5.5 mg/l (nominal conc.), respectively. Thus, based on the LC50 value, chemical was considered as toxic to aquatic fish. Since, the test chemical is readily biodegradable in water, chemical was considered to be non-toxic to aquatic fish at environmental relevant concentrations and thus considered to be 'not classified' 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 in fish. Study was performed following the EU Method C.1 (Acute Toxicity for Fish) (Council Directive 92/69/EEC C.1 (1992)). Danio rerio (Zebra fish) obtained from west-aquarium of 2.5 to 3.5 cm length was used as a test organism for the study. Test chemical concentrations were verified analytically by HPLC at the start and after 24 hr of the experiment. Nominal test chemical concentrations used for the study were 0, 2.8, 3.9, 5.5 and 7.8 mg/l, respectively. The test solution was preared by dissolved the test chemical into synthetic water and treated for 60 seconds at 8000 rpm with an ultra-turrax. Test fishes were exposed with the test chemical in a close ventilated test vessel. Study was performed using test fishes in a semi-static system at 20.7-22.8°C temperature, 7.5 - 8.2 pH, 8.2 - 9.6 (mg/l) dissolved oxygen and hardness 14.5 [°dH] under a photoperiod of 12:12 light:dark conditions for 96 hrs. Daily renewal of the test medium was done during the study. The LC0 and the LC100 are determined directly from the study, while the LC50 is calculated or read from the dose-response curve. On the basis of the effect of test chemical on the mortality of the test organism, the 24hr LC0, LC50 and LC100 value was determined to be 1.41 mg/l (measured conc.), 2.35 mg/l (measured conc.) & 3.93 mg/l (measured conc.) and the 96 hr LC50 & LC100 value was determined to be in the range of > 3.9 to < 5.5 mg/l (nominal conc.) and 5.5 mg/l (nominal conc.), respectively. Thus, based on the LC50 value, chemical was considered as toxic to aquatic fish. Since, the test chemical is readily biodegradable in water, chemical was considered to be non-toxic to aquatic fish at environmental relevant concentrations and thus considered to be 'not classified' as per the CLP classification criteria.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

3.9 mg/L

Additional information

Various experimental studies of the test chemical were reviewed for the short term toxicity to fish end point which are summarized as below:

 

An acute toxicity test was conducted for 96 hrs for assessing the effect of test chemical in fish. Study was performed following the EU Method C.1 (Acute Toxicity for Fish) (Council Directive 92/69/EEC C.1 (1992)). Danio rerio (Zebra fish) obtained from west-aquarium of 2.5 to 3.5 cm length was used as a test organism for the study. Test chemical concentrations were verified analytically by HPLC at the start and after 24 hr of the experiment. Nominal test chemical concentrations used for the study were 0, 2.8, 3.9, 5.5 and 7.8 mg/l, respectively. The test solution was preared by dissolved the test chemical into synthetic water and treated for 60 seconds at 8000 rpm with an ultra-turrax. Test fishes were exposed with the test chemical in a close ventilated test vessel. Study was performed using test fishes in a semi-static system at 20.7-22.8°C temperature, 7.5 - 8.2 pH, 8.2 - 9.6 (mg/l) dissolved oxygen and hardness 14.5 [°dH] under a photoperiod of 12:12 light:dark conditions for 96 hrs. Daily renewal of the test medium was done during the study. The LC0 and the LC100 are determined directly from the study, while the LC50 is calculated or read from the dose-response curve. On the basis of the effect of test chemical on the mortality of the test organism, the 24hr LC0, LC50 and LC100 value was determined to be 1.41 mg/l (measured conc.), 2.35 mg/l (measured conc.) & 3.93 mg/l (measured conc.) and the 96 hr LC50 & LC100 value was determined to be in the range of > 3.9 to < 5.5 mg/l (nominal conc.) and 5.5 mg/l (nominal conc.), respectively. Thus, based on the LC50 value, chemical was considered as toxic to aquatic fish. Since, the test chemical is readily biodegradable in water, chemical was considered to be non-toxic to aquatic fish at environmental relevant concentrations and thus considered to be 'not classified' as per the CLP classification criteria.

 

Another short term toxicity to fish study was conducted for 96 hrs for assessing the effect of test chemical (Experimental study report, 2013). The test was performed in accordance to OECD guideline No. 203 “Fish Acute Toxicity Test”. Poecilia reticulata (Guppy fish) of age 3 -4 months old, average weight 0.25 g and average length of 2.0 to 2.5 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. The test conditions during the housing of the test organisms were oxygen content of 7.4 mg/l, pH 7 -8, water temperature 25°C and under a photoperiod of 12:12 hr light: dark conditions, respectively. The test solution was prepared by dissolving 15mg, 25mg and 35mg of the test substance in 10 liters deionized water separately with continuous stirring for achieving the test concentration of 1.5 mg/L, 2.5 mg/L & 3.5 mg/L, respectively. Range finding test was conducted with the several test concentrations, i.e., 1.5 mg/L, 2.5 mg/L, 3.5 mg/L, 6.5 mg/L, 25 mg/L & 50 mg/L. But at 6.5 mg/L, 25 mg/L & 50 mg/L 100% mortality was observed within 24 hrs. Therefore, the experiment was continued only for three concentrations, i.e., 1.5 mg/L, 2.5 mg/L, 3.5 mg/L in which no mortality was found. In definitive study, limit test was conducted using a 100 mg/l test chemical concentration. Total 8 fishes were exposed to test chemical in a 5 lit bowl aquaria containing 4 liters of potable water. The test vessels were placed under a photoperiod of 12:12 hr light: dark conditions for 96 hrs. Aeration in test vessels was provided 1 day before the start of the experiment. The fishes were moving slowly in the test chemical conc. as compared to the control. No mortalities were observed in the control and test vessel during the range finding study (at all three test concentrations i.e; 1.5, 2.5 and 3.5 mg/l). On the basis of this, the 96 hr LC0, median lethal concentrations (LC50) and LC100 value for test chemical on Poecilia reticulata (Guppy fish) was determined to be 3.5 mg/l, > 3.5 to < 6.5 mg/L and 6.5 mg/l, respectively.

 

For the test chemical, short term toxicity to fish study was conducted for 96 hrs for assessing the effect of test chemical (Secondary source, 2019). Lepomis macrochirus (Bluegillfish) was used as a test organism. Study was performed in a static system. On the basis of the effect on mortality of the test organism, the 96 hr NOEL and LC50 was determined to be 1.97 and > 20 mg/l. Thus, test chemical was considered as non-toxic to aquatic fish.

 

Additional short term toxicity to fish study from secondary source (2019) was conducted for 96 hrs for assessing the effect of test chemical. Lepomis macrochirus (Bluegillfish) was used as a test organism. Study was performed in a static system. On the basis of the effect on mortality of the test organism, the 96 hr NOEL and LC50 was determined to be 0.7 and 2.99 mg/l (95% C. I. = 2.24 to 4.0 mg/l), respectively.

 

Another short term toxicity to fish study was conducted for 96 hrs for assessing the effect of test chemical (2019). Oncorhynchus mykiss (Rainbow Trout) was used as a test organism. Study was performed in a static system. On the basis of the effect on mortality of the test organism, the 96 hr NOEL and LC50 was determined to be 1.25 and 1.67 mg/l (95% C. I. = 1.25 to 2.24 mg/l), respectively..

 

In a supporting weight of evidence study from review article and secondary source, short term fish toxicity was conducted for 47 hrs for assessing the effect of test chemical. Cyprinus carpio (Common carp) captured with an alternating-current electric boat shocker in the New York State Barge Canal of weight 1-10 pounds (average weight of around 3 pounds) was used as a test organism for the study. After transportation of test organism to the laboratory, oxygen was supplied. Test fishes were acclimatized for weeks at spring temperature of 47°F. Test chemical concentrations were not verified analytically. Test chemical concentrations used for the study were 81 and 163 mg/l (nominal concentrations), respectively. Study was performed using fishes in a flow through system at 18.33°C (65°F) temperature, 6.7 pH and alkalinity of 10 mg/l CaCO3. Test fishes were exposed to test chemical in 350 and 550 gallons glass-fronted, fiberglass tank for 47 hr. After exposure period of 47 hr, mortality and other visual symptoms of the test organism was noted. No abnormal responses of the test fish were observed. On the basis of the effect on mortality of the test organism Cyprinus carpio, the 47 hr NOEC was determined to be in the range of 81-163 mg/l (nominal concentration).

 

On the basis of the above results, it can be concluded that the test chemical was consideredastoxic to fish.Since, the test chemical is readily biodegradable in water, chemical was considered to be non-toxic to aquatic fish at environmental relevant concentrations and hence, considered to be 'not classified' as per the CLP classification criteria.