3-ethoxy-4-hydroxybenzaldehyde

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

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 without detailed documentation

Remarks:

The test conditions are well described and are in accordance with the recognised EU and OECD test guidelines. The GLP are not mentioned.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

yes

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: Toxicants were proportionally diluted with lake superior water from stock solution before delivery to fish exposure chamber.
Test chemicals were prepared for proportional dilution by the diluted stock method. i e : premixed aqueous solution of desired concentration.
Stock solution: 1000 mg/L
- Controls: blank

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Pimephales promelas (Fathead minnow)
- Source: From US EPA Environmental Research Laboratory- Duluth
- Age at study initiation: 28 days
- Weight at study initiation: Mean: 0.034 g; SD: 0.0131 g
- Length at study initiation: Mean: 14.3 mm; SD: 2.245 mm
- Method of breeding: Adults were held at 25°C in flowing water with a 16-h light-controlled photoperiod and fed frozen adult brine shrimp. They were provided with asbestos pipe (cut in half longitudinally) as spawning substrate. Eggs were spawed and fertilized on the undersides of these substrates. The embryos were tended by the male parent until eye-up at which time the substrates with embryos attached were removed to another 25°C bath where hatching occured.
- Feeding during test: no

ACCLIMATION
- Acclimation period: no

QUARANTINE: no data

Test type:

flow-through

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

behaviour also reported during the test

Hardness:

52.2 mg/L CaCO3 (SD: 0.65) measured by the compleximetric or EDTA method

Test temperature:

24.1 °C (SD: 0.83) measured daily using partial immersion of a mercury thermometer

pH:

7.69 (SD: 0.28) measured with a meter calibrated shortly prior each test

Dissolved oxygen:

6.5 mg/L (SD: 0.41) measured with an oxygen sensitive electrode calibrated weekly using the azide modification of the winkley method

Salinity:

no data (fresh water)

Nominal and measured concentrations:

nominal concentration : 0 / 12.5 / 25 / 50 / 100 / 200 /
measured concentrations : see table 1 in "any other information"

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 3.4 l glass open tank
- Aeration: no
- Type of flow-through: cycling proportional diluters
- Renewal rate of test solution: no data
- No. of organisms per vessel: 22 to 26 fish per tank
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 0

WATER PARAMETERS:
- Source: Lake Superior water
- Total organic carbon:
- Metals: Al (1-26 µg/L); Cd (<0.1 µg/L); Cr (2-20 µg/L); Co (<0.5 µg/L); Cu (0.3-3.2 µg/L); Fe (2-83 µg/L); Pb (7-20 µg/L); Mn (0.2-11.5 µg/L); Ni (<0.5 µg/L); Zn (1-2.7 µg/L)
- Intervals of water quality measurement: at least once for each test.

TEST MEDIUM PARAMETERS:
- Test temperature: 24.1 °C
- dissolved oxygen: 6.5 mg/l
- hardness (CaCO3): 52.5 mg/L
- pH: 7.69

OTHER TEST CONDITIONS: no data
photoperiod : no data

EFFECT PARAMETERS MEASURED: mortality

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study
- Test concentrations: 0, 12.5, 25, 50, 100 and 200 mg/L

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

87.6 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 81.4 - 94.3

Details on results:

Affected fish swan at surface. Respiration and gill movements were limited. Fish were also hypoactive and lost equilibrium prior to death.

Results with reference substance (positive control):

not relevant

Reported statistics and error estimates:

no data

Sublethal observations / clinical signs:

Table 2: Mortalities observed in each exposure chamber

Hour	control		A		B		C		D		E
Initial n° of fish in each exposure chamber	26	25	25	25	23	22	26	24	25	26	25	25
1.75	1	0	0	1	1	0	1	0	0	1	3	2
3	1	0	0	1	1	0	1	0	0	1	3	4
6	1	0	0	1	1	0	1	0	0	1	4	6
12	1	0	0	1	1	0	2	0	0	1	21	20
24	1	0	0	1	2	0	3	0	0	1	22	22
29.5	1	0	0	1	2	0	3	0	0	1	22	22
48	1	0	0	1	2	0	3	0	3	1	24	24
53.5	1	0	0	1	2	0	3	0	3	1	24	24
72	1	0	0	1	2	0	3	0	3	1	25	25
77.2	1	0	0	1	2	0	3	0	3	1	25	25
96	1	0	0	1	2	1	3	0	3	1	25	25

Validity criteria fulfilled:

not specified

Conclusions:

With an EC50-96h = 87.6 mg/L for Pimephales promelas, ethylvanillin is harmful for fish.
The validity criteria were not examined by the authors but 3 of the 4 validity criteria detailled in 203 OECD guideline were fullfilled:
- mortality in the controls did not exceed 10% at the end of the test
- the test was conducted under flow through conditions
- deviation of measured concentration from nominal ones was greater than 20% BUT the results were based on measured concentrations.

Concerning oxygen concentration, it is described that "test mean concentration were maintained between 60 and 100 % of water saturation except in 12 tests" it is not clear if it is the case for ethylvanillin test.

Executive summary:

In a 96-h acute toxicity study (Brooke, 1984), Fathead Minnows (Pimephales promelas) were exposed to ethyl vanillin at nominal concentrations of 0 (control); 12.5; 25; 50; 100 & 200 mg/L under flow through conditions. 

The 96-h LC₅₀ was 87.6 mg a.i./L.  95% C.I.:  81.4 to 94.3 mg/L (based on measured concentrations).

Based on the criteria of the CLP regulation, Ethylvanillin is considered as harmful for Pimephales promelas in the conditions tested.

Although GLP were not mentionned and validity criteria not detailled, this toxicity study satisfies the guideline requirement and is therefore considered as reliable with restrictions.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1963

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Study well perfomed even if not GLP (1963). Only 3 animals tested with a force feeding method far from recommended guidelines.

Principles of method if other than guideline:

Test material were forced fed to fish and behaviour and death were recorded during a minimum of 40h.
1,496 chemicals were tested including Ethyl vanillin

GLP compliance:

no

Analytical monitoring:

no

Test organisms (species):

Cyprinus carpio

Details on test organisms:

- capturation: with an alternating-current electric boat shoker in the New York State Barge Canal.
- Transportation to the lab by tank truck supplied with oxygen
- Acclimation for weeks at a temperature of 47°F (i.e. 8.3°C)
- weight: form 1 to 10 pounds average = 3 pounds (i.e. from 0.5 to 5 kg with a mean at 1.5 kg)
- poor quality fish were rejected
- fish were maintained into 350 to 550 L fiberglass tanks under a clear, continously flowing spring (50 to 100 L/min)
- water quality: pH 6.7 with 10 ppm of alkalinity and traces of other compounds and elements
- feeding during test: yes

no more data

Test type:

other: force feeding

Water media type:

freshwater

Limit test:

no

Total exposure duration:

44 h

Test temperature:

18.3°C (67°F)

pH:

6.7

Dissolved oxygen:

no data

Nominal and measured concentrations:

Nominal: 205, 212 and 222 mg/L

Details on test conditions:

3 fish were tested at 205, 212 and 222 mg/L
Test substances were placed in one or two n°5 gelatin capsules.
Dissection showed that gelatin capsules in these conditions of 18.3°C disintegrated in approximatively one hour.
The 3 fish were removed from 8.3°C water, force-fed, tagged, and placed in 18.3°C running water for observation.
They quickly adjust to this temperature and sometimes attempted to feed immediatly

Reference substance (positive control):

not specified

Key result

Duration:

44 h

Dose descriptor:

LC0

Effect conc.:

205 - 222 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

1 h has been deducted from exposure time of 44h.
It corresponds to the disintigration time of the gelatin capsule containing the test substance in the test conditions.

Validity criteria fulfilled:

not applicable

Conclusions:

Ethyl vanillin should not be toxic to carps after ingestion of 205 to 222 mg/L

Executive summary:

Ethylvanillin was forced fed to fish and behaviour and death were recorded during a minimum of 44 hours.

The administration of 205 to 222 mg/L do not induce any toxicity to the fishes.

Ethylvanillin is not considered harmful to fish, under these conditions.

Description of key information

The 96-hour LC50 of test item to Pimephales promelas was determined to be 87.6 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

87.6 mg/L

Additional information

Two studies were assigned a Klimisch score of 2. One of them, done in accordance with the recognized guidelines, was selected as key study, and its summary is the following:

In a 96-h acute toxicity study (Brooke, 1984), fathead minnows (Pimephales promelas) were exposed to ethylvanillin at nominal concentrations of 0 (control); 12.5; 25; 50; 100 and 200 mg/L under flow through conditions. 

The 96-h LC₅₀ was 87.6 mg/L.  95% C.I. (Confidence Interval):  81.4 to 94.3 mg/L (based on measured concentrations).

Based on the criteria of the CLP regulation, ethylvanillin is considered as harmful for Pimephales promelas.

Although GLP were not mentioned and validity criteria not detailed, this toxicity study satisfies the guideline requirement and is therefore considered as reliable with restrictions.