Benzenepropanoic acid, .beta.,4-dimethoxy-, methyl ester

Short-term toxicity to fish

Administrative data

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:

other: Guideline study conducted in accordance with GLP, however the test substance had a very low purity (e.g. < 90 %).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

Cited as Directive 84/449/EEC, C.1 ("Acute toxicity for fish")

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

methyl 3-methoxy-3-(4-methoxyphenyl)propanoate

EC Number:

614-925-8

Cas Number:

69098-08-4

Molecular formula:

C12 H16 O4

IUPAC Name:

methyl 3-methoxy-3-(4-methoxyphenyl)propanoate

Details on test material:

- Name of test material (as cited in study report): Benzenepropanoic acid. -beta-, 4-dimethoxy-, methyl ester, technical grade (3-(4-Methoxyphenyl)-3-methoxypropionsaeuremethylester
techn.)
- Lot/batch No.: 165
- Analytical purity: 71.2 %
Water-solubility: slight

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Due to the insolubility of the test substance in water, 100mg/l of Tween 80 was added to the test solutions, acting as a solubiliser. The highest test concentration (100 mg/l) was tested twice, once with addition of Tween 80 and once without. After liquefying the test material 22.8, 48.7, 104.5. 226.3, 487.7 and 1051 mg test substance were each separately mixed with 1 g Tween 80 (0.1 g/l) for each test concentration. Each of these blends were further mixed with small quantities of bidistilled water to prepare a dispersion and then transferred into the test water using an ultra-turrax stirrer. Analogously 1 g Tween 80 was added to a carrier control. 1050.1 mg test substance without Tween 80 was added to the test water in the same way. Subsequently the fish were placed into the aquaria within 25 min. after the test material was added.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: Zebra fish
- Age at study initiation (mean and range, SD): about 5 months
- Length at study initiation (length definition, mean, range and SD): 3.29 cm (range: 2.8 - 3.6 cm)
- Weight at study initiation (mean and range, SD): 0.305 g (range: 0.16 - 0.45 g)
- Corpulence factor of the batch: 0.86
- Feeding during test: 1 day before and during exposure

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

approx. 2.5 mmol/l

Test temperature:

22 - 24 °C (temperature-controlled room)

Nominal and measured concentrations:

Nominal test concentrations: 2.15, 4.64, 10.0. 21.5, 46.4 and 100 mg/l and an untreated control.

Details on test conditions:

TEST SYSTEM
- Fill volume: 10 l
- Aeration: none
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.3 g fish / l test water


TEST MEDIUM / WATER PARAMETERS
- Acid capacity: approx. 5.5 mmol/l


OTHER TEST CONDITIONS
- Photoperiod: 16 : 8 hours day-night regime


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and symptoms of intoxication

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.2

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

13.1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

13.1 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 17.4 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 4

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

43.8 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Reported statistics and error estimates:

Determination or calculation of the median of the LC50 and if possible the LC5 and the LC95 using the probit analysis after (nominal conc.) 1, 4, 24, 48, 72, 96 h.

Any other information on results incl. tables

Sublethal observations / clinical signs:

During the 96 h exposure to 3-(4-Methoxyphenyl)-3-methoxypropionsaeuremethylester
techn., mortalities were as follows:

Nominal test Cumulative mortalitiy (%) after x hours
concentration 1 / 4 / 24 / 48 / 72 / 96
(mg/l)
Control 0 / 0 / 0 / 0 / 0 / 0 /
Control* 0 / 0 / 0 / 0 / 0 / 0 /
2.15 0 / 0 / 0 / 0 / 0 / 0 /
4.64 0 / 0 / 0 / 0 / 0 / 0 /
10.0 0 / 0 / 0 / 0 / 0 / 0 /
21.5 0 / 0 / 3 / 5 / 6 / 7 /
46.4 0 / 8 / 10 / 10 / 10 / 10 /
100 1 / 10 / 10 / 10 / 10 / 10 /

100** 1 / 10 / 10 / 10 / 10 / 10 /

* = solvent control
** = highest test concentarion without Tween 80

Symptoms of intoxication:
Apathy, a narcotic-like state and tumbling were observed within test concentration 21.5 mg/l (nominal) and upwards.
The results referring to the highest tested concentration of 100 mg/l with or without addition of Tween 80 were similar.

Nominal test Symptoms of intoxication after x hours
concentration 1 / 4 / 24 / 48 / 72 / 96
(mg/l)
Control - / - / - / - / - / - /
Control* - / - / - / - / - / - /
2.15 - / - / - / - / - / - /
4.64 - / - / - / - / - / - /
10.0 - / - / - / - / - / - /
21.5 - / AT / A / A / A / A /
46.4 ANT / N / - / - / - / - /
100 NT / - / - / - / - / - /

100** N / - / - / - / - / - /

Definitions of Symptoms:
-: No symptoms
+: All fish dead
A: Apathy
N: Narcotic-like state
T: Tumbling

On the basis of the nominal concentrations after 96 h:
LC 50: approx. 19 mg/l
NOEC: 10.0 mg/l
LC0: 0.0 mg/l
LC100: 46.4 mg/l
--------------------------------------------------------
On the basis of the analytically measured concentrations after 96 h:
LC 50: approx.15 mg/l
NOEC: 10.7 mg/l
LC0: 10.7 mg/l
LC100: 40.3 mg/l
--------------------------------------------------------
On the basis of the analytically measured concentrations after 1 and 96 h:
Values after 1 and 96 hours
Median LC 50: approx. 17.4 mg/l
Median NOEC: 13.1 mg/l
Median LC0: 13.1 mg/l
Median LC100: 43.8 mg/l
--------------------------------------------------------

Analytical results:
The analytically detected concentrations were initially in the range of 95.3 % - 200.4 % and after about 96 hours between 77.2 % - 209.3 % of the nominal concentrations.