Reaction mass of (3R)-3-[(1R)-1-ethoxyethoxy]-3-methylpent-1-en-4-yne and (3R)-3-[(1S)-1-ethoxyethoxy]-3-methylpent-1-en-4-yne and (3S)-3-[(1R)-1-ethoxyethoxy]-3-methylpent-1-en-4-yne and (3S)-3-[(1S)-1-ethoxyethoxy]-3-methylpent-1-en-4-yne

Short-term toxicity to fish

Administrative data

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:

other: Guideline study in compliance with GLP-criteria.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1998

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

72008-25-4

EC Number:

615-703-3

Cas Number:

72008-25-4

IUPAC Name:

72008-25-4

Details on test material:

- Name of test material (as cited in study report): Asta-C6-Acetal rein
- Physical state: liquid, yellowish
- Homogeneity: homogeneous
- Storage: room temperature, exclusion of light.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples taken just before test solution renewal (at ca. 24h, 48h 72h and 96h) and at teh start of the test (ca. 1 h).

Test solutions

Vehicle:

no

Details on test solutions:

For the first substance preparation (t=0), to obtain test concentrations of 4.64, 10, 21.5 and 46.4 mg/L, test substance at 0.0465, 0.1001, 0.2152, 0.4641 and 1.00 g were added to the test water using an ultra-turrax stirrer.
For the second substance preparation (t=24h), to obtain test concentrations of 4.64, 10, 21.5 and 46.4 mg/L, test substance at 0.0465, 0.1004, 0.2153 and 0.4644 g were added to the test water using an ultra-turrax stirrer.
For the third substance preparation (t=48h), to obtain test concentrations of 4.64, 10, 21.5 and 46.4 mg/L, test substance at 0.0464, 0.1001, 0.2151 and 0.4640 g were added to the test water using an ultra-turrax stirrer.
For the fourth substance preparation (t=72h), to obtain test concentrations of 4.64, 10, 21.5 and 46.4 mg/L, test substance at 0.0462, 0.1006, 0.2151 and 0.4642 g were added to the test water using an ultra-turrax stirrer.
From the beginning of preparing the test solutions until termination of placing the fish into the aquaria 20 minutes were needed.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

GENERAL INFORMATION:
- Animal Species: Zebra fish (Brachydanio rerio RAM. and BUCH.)
- Consignment dated: Jun. 24, 1997
- Animal. breeder/suppijer: Charles River Aquatics, Someren; Netherlands
- Hatching date: April 03, 1997
- Age of the animals: About 3 months
- Body length: 3.67 (3.2 - 3.9) cm
- Body weight: 0.55 (0.3 - 0.8) g

HOUSING:
- The fish were kept in a flow-through tank in tap water, not chlorinated, passed through a charcoil. filter and aerated with oil-free air.
-Duration of housing: About 2 months
- Photoperiod: 16 hours light and 8 hours dark
- Total hardness: About 2.5 mmol/L (= about 250 mg/L CaCO3)
- Acid capacity: About 5.5 mmol/L
- Oxygen content: > 60% of maximum saturation
- pH: About 8.0
- Water temperature: About 22 - 23 °C
- Mortality during the last 14 days: 0 %
- Medical treatment: none
- Diet: 'TETRA MIN' standard feed for aquarium fish (TETRA - WERKE, MELLE, FRG) ad libitum; and on workdays generally additionally live artemia.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Test conditions

Hardness:

2.5 mmol/L (about 250 mg/L CaCO3)

Test temperature:

22 - 23 °C

pH:

8.1-8.5

Dissolved oxygen:

5.4-8.5 mg/L

Nominal and measured concentrations:

Nominal concentrations:
0 (control), 4.64, 10, 21.5, 46.4 and 100 mg/L.
Measured concentrations (just before test medium renewal):
- After 24h: n.d., 3.1, 6.5, 16.1, 32.8 and 75.2 mg/L.
- After 48 h: n.d., 2.4, 6.5, 14.9 and 35.6 mg/L (nominal conc. of 100 mg/L not analysed).
- After 96h: n.d., 3.0, 7.2, 15.4 and 36.8 mg/L (nominal conc. of 100 mg/L not analysed).

Details on test conditions:

TEST SYSTEM
- Test vessel: 10-L all-glass aquaria, open, 30 x 22 x 24 cm
- Aeration: aeration with oil-free air
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.6 g fish /L

TEST MEDIUM / WATER PARAMETERS
Municipal water of the City of Frankenthal, not chlorinated and passed through a charcoal filter, aerated

OTHER TEST CONDITIONS
- Photoperiod: 16:8 hour light:dark
- Adaptation: Test conditions corresponded to housing conditions.
- Withdrawal of food: 1 day before and during exposure.

EXAMINATIONS:
- Mortality: at 1, 24, 48, 72, and 96 hours
- Symptoms: apathy, abdominal distension, exophtalmos, escape reflex, hyperreflexia, convulsions, gasping, narcotic-like state, tumbling, restlessness, discoloration, headstand and accelerated respiration at 1, 24, 48, 72, and 96 hours
- the analytical concentrations after 1, 24, 48, 96 and 1+24+48+96 hours.

Reference substance (positive control):

not specified

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

21 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.L.: 14.3-32.6 mg/L

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

27.35

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% C.L.: 19.46-38.22 mg/L

Details on results:

Mortality occurred first at the nominal concentration 21.5 mg/L.
Other substance-related effects were: apathy, convulsians, narcotic-like state, tumbling, swimming near water surface, swimming near the bottom.

Reported statistics and error estimates:

Finney, D.J., Probit Analysis, Cambr. Univ. Press, 3rd ed., 1971; certain aspects of this method have been modified. The exact procedure can be
seen from the Standard Operating Procedures (SOPs) of the laboratory.

Any other information on results incl. tables

Sublethal observations / clinical signs:

Mortality:

Nominal conc. (mg/L)	Number of fish	Mortality (cumulated) after
		1h	4h	24h	48h	72h	96h
0	10	0	0	0	0	0	0
4.64	10	0	0	0	0	0	0
10	10	0	0	0	0	0	0
21.5	10	0	0	0	0	1	4
46.4	10	0	0	2	3	7	8
100	10	7	10	10	10	10	10

Analytically determined concentrations:

Nominal conc. (mg/L)	Measured conc. (mg/L) and recovery (%) after:				Aritmetic mean measured conc. (mg/L)
	1h	24h	48h	96h	1h+24h+48h+96h
0	n.d.	n.d.	n.d.	n.d.	-
4.64	3.6 (77.6%)	3.1 (66.8%)	2.4 (51.7%)	3.0 (64.7%)	3.0
10	8.6 (86.0%)	6.5 (65.0%)	6.5 (65.0%)	7.2 (72.0%)	7.2
21.5	18.9 (87.9%)	16.1 (74.9%)	14.9 (69.3%)	15.4 71.6%)	16.3
46.4	41.0 (88.4%)	32.8 (70.7%)	35.6 (76.7%)	36.8 (79.3%)	36.5
100	91.4 (91.4%)	75.2 (75.2%)	n.a. -	n.a. -	83.3

n.d. = not detectable

n.a. = not analysed