Reaction mass of (2R,7aR)-2,4,6,6-Tetramethyl-7,7a-dihydro-1,3-benzodioxol-5(6H)-one and (2S,7aS)-2,4,6,6-Tetramethyl-7,7a-dihydro-1,3-benzodioxol-5(6H)-one

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:

162691-58-9

EC Number:

605-300-0

Cas Number:

162691-58-9

IUPAC Name:

162691-58-9

Details on test material:

- Name of test material (as cited in study report): Asta-C9-Acetal
- Physical state: oily, yellowish
- Homogeneity: on visual inspection homogeneous
- Storage consitions: room temperature

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 the start of the test (ca. 1 h).

Test solutions

Vehicle:

yes

Details on test solutions:

For the first substance preparation (t=0), to obtain test concentrations of 4.64, 10.0, 21.5, 46.4 and 100 mg/L, test substance at 0.0463, 0.1001, 0.2151, 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, 10.0, 21.5 and 46.4 mg/L, test substance at 0.0463, 0.1004, 0 .2151 and 0 .4639 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, 10.0, 21.5 and 46.4 mg/L, test substance at 0.0465, 0.1001, 0.2152 and 0.4641 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, 10.0, 21.5 and 46.4 mg/L, test substance at 0.0466, 0.1001, 0.2151 and 0.4644 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.

* No 100 mg/L test solution prepared as cumulative mortality reached 100% already at 24h after exposure.

Test organisms

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST SPECIES:
- Animal Species: Zebra fish (Brachydanio rerio HAM . and BUCH .)
- Consignment dated: Jun. 24, 1997
- Animal breeder/supplier: Charles River Aquatics, Someren, Netherlands
- Hatching date: April 03, 1997
- Age of the animals: About 4 months
- Body length: 3.8 (3.4 - 4.2) cm
- Body weight: 0.47 (0.27 - 0.67) g

HOUSING
- The fish were kept in a flow-through tank in tap water, not chlorinated, passed through a charcoal 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 CaC03)
- Acid capacity: about 5.5 mmol/L
- Oxygen content: > 60% of maximum saturation
- pH: About 8 .0
- Water temperature: about 21-23 °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.

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

Test temperature:

22 - 23 °C (temperature-controlled room)

pH:

7.9-8.5

Dissolved oxygen:

5.6 - 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, 5.5, 15.2, 33.5 and 71.2 mg/L.
- After 48 h: n.d., 3.2, 7.2, 15.5 and 33.8 mg/L (nominal conc. of 100 mg/L not analysed).
- After 96h: n.d., 3.3, 7.2, 15.9 and 34.7 mg/L (nominal conc. of 100 mg/L not analysed).

Details on test conditions:

TEST SYSTEM
- Test vessel: 10-L all-glass aquaria (one per test group), open, 30 x 22 x 24 cm, about 17 cm water depth.
- 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: 0.5 g fish /L

TEST WATER:
- Municipal water of the city of Frankenthal, not chlorinated and passed through a charcoal filter, aerated. The test water is regularly assayed for chemical contaminants by the municipal authorities of Frankenthal and the technical services of BASF Aktiengesellschaft as well as for presence of microbes by a contract laboratory.

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.

EFFECT PARAMETERS MEASURED:
- 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.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

54.9 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: Mean values of the analytically detected concentrations after 1+24+48+72+96 hours. The LC50 value was calculated as the geometrical mean of the LC0 and LC100 nominal values.

Effect concentrations 2

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

68.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: The LC50 value was calculated as the geometrical mean of the LC0 and LC100 nominal values.

Details on results:

- Mortality: deaths were reported only at the highest tested concentration of 100 mg/L. First deaths occurred after 4 hours exposure (3/10), after 24 hours all fish had died (i.e. cumulated mortality 100%).
- Other substance-related effects: apathy, narcotic-like state, tumbling, swimming near water surface and 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.

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
10.0	10	0	0	0	0	0	0
21.5	10	0	0	0	0	0	0
46.4	10	0	0	0	0	0	0
100	10	0	3	10	10	10	10

Analytically determined concentrations:

Nominal conc. (mg/L)	Measured concentration after:
	1h	24h	48h	72h	96h
0	n.d.	n.d.	n.d.	n.d.	n.d.
4.64	4.2 (90.5%)	3.1 (66.8%)	3.2 (69.0%)	3.1 (66.8%)	3.3 (71.1%)
10.0	7.6 (76.0%)	5.5 (55.0%)	7.2 (72.0%)	7.0 (70.0%)	7.2 (72%)
21.5	19.4 (90.2%)	15.2 (70.7%)	15.5 (72.1%)	15.4 (71.6%)	15.9 (74.0%)
46.4	45 (97.0%)	33.5 (72.2%)	33.8 (72.8%)	34.5 (74.4%)	34.7 (74.8%)
100	95 (95.0%)	71.2 (71.2%)	n.a. -	n.a. -	n.a. -

n.d. = not detectable

n.a. = not analysed