Registration Dossier

Administrative data

Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
November 17 to november 28, 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: OECD, GLP study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report date:
1998

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
GLP compliance:
yes
Remarks:
Principles of Good Laboratory Practice, annex of paragraph 19a, section 1 of the chemical law.

Test material

Constituent 1
Reference substance name:
Azacyclonol
EC Number:
204-092-5
EC Name:
Azacyclonol
Cas Number:
115-46-8
IUPAC Name:
diphenyl(piperidin-4-yl)methanol
Test material form:
solid - liquid: suspension
Details on test material:
- Name of test material (as cited in study report): Azacyclonol-Base
- Physical state: white powder
- Purity: 98.3 % (w/w)
- Batch No.: 86000298
- Storage conditions: darkness at approximately 5 °C in a refrigerator
- Date of expiry: September 10, 1998
- Solubility in water: not soluble

Sampling and analysis

Analytical monitoring:
yes
Details on sampling:
Aqueous solutions of 20ml diluted in methanol.

Test organisms

Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Zebra fish
- Strain: Danio rerio
- Origin: Hoechst Marion Roussel Deutschland GmbH Global Preclinical Development - Drug Safety
- Length at study initiation: 2.6 cm ± 0.08 (variation range: 2.5-2.7)
- Food type: Tetra Min, Tetra Werke, Melle (Germany)

ACCLIMATION
- Acclimation period: 14 days in water for dilution
- Acclimation conditions: light period: 12 hours daily
- Population density: <= 1 g fish/L water
- Food type:Tetra Min, Tetra Werke, Melle (Germany)
- Feeding frequency: twice daily ad libitum

Study design

Test type:
static
Water media type:
not specified
Total exposure duration:
96 h

Test conditions

Hardness:
In a range of 2.3 - 2.5 mmol Ca2+ + Mg2+/ L during the study.

Test temperature:
22 .± 1 °C
pH:
Initial pH in study batch: in the range of 8.3-8.5
pH after adding the substance in the 500 mg/L group: 9.6. The pH was adusted with concentrated HCI.
pH after substance addition in the 100 and 220 mg/L groups was below the maximum tolerable value of 9.
Dissolved oxygen:
>= 80% of the saturation value
Oxygen content in concentration groups: 7.0 - 8.8 mg/L
Nominal and measured concentrations:
Test concentrations: 0, 100, 220 and 500 mg/L
Nominal concentrations: 0, 100 and 220 mg/L
Measured concentrations: 111 and 243 mg/L
Details on test conditions:
TEST SYSTEM
- Test vessel: test chambers, which were calibrated to 4 liters
- Aeration: test chambers were not aerated during the course of the study.
- No. of organisms per vessel: 7 fishes
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 1

OTHER TEST CONDITIONS
- Adjustment of pH: with HCl
- Photoperiod: from above from 06.00 a.m to 06.00 p.m
- Light intensity: 700 lux

TEST CONCENTRATIONS
Experimental part 1: 7 fish allocated in 2 chambers received 0 and 500 mg/L.
Experimental part 1: 7 fish allocated in 3 chambers received 0, 100 and 220 mg/L.

Results and discussion

Effect concentrationsopen allclose all
Duration:
96 h
Dose descriptor:
LC0
Effect conc.:
ca. 100 mg/L
Nominal / measured:
nominal
Remarks on result:
other: With the exception of the value observed at study start in the 100 mg/L group (123% of the nominal concentration) all values determined were in a range of± 20 % of the theoretical value.
Duration:
96 h
Dose descriptor:
LC50
Effect conc.:
ca. 148 mg/L
Nominal / measured:
meas. (not specified)
Basis for effect:
mortality (fish)
Duration:
96 h
Dose descriptor:
LC100
Effect conc.:
ca. 220 mg/L
Nominal / measured:
nominal
Basis for effect:
mortality (fish)
Remarks on result:
other: All values determined were in a range of ± 20 % of the theoretical value.
Details on results:
In all tested concentration groups: changes in appearance, behavior, swimming behavior and respiration rate during the entire exposure time.
ln the 500 mg/L groups: recording of symptoms was not possible because of 100 % lethality at the first inspection date.
ln the 2200 mg/L group: symptoms and lethality were observed.
ln the 100 mg/L group: symptoms but no lethality.
Dead fish showed substance deposits on integument, metallic sheen or dark coloured integument.
Reported statistics and error estimates:
Probit analysis could not be carried out because of the lethality rate. As the factor for the concentration steps was 2.2 (third root of 10), no further concentrations were tested.The geometrie mean of the LC0 and LC100 is given.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The LC50 of Azacyclonol-Base after 96 hours was 148 mg/L.
Azacyclonol base induced an increase of the pH > 9 in the test solutions; therefore pH was adjusted by adding concentrated HCl. The ecotoxicity profile of free base could be used as reference.