Ichthammol

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Start of study: February 17th, 1995
Termination of study: March 24th, 1995

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

The study was carried out according to the OECD Method No. 203 dated April 4th, 1984 and EC guideline C.l. published in the Official Journal of the European Communities L 383 A dated December 29lh, 1992.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

Ichthammol

EC Number:

232-439-0

EC Name:

Ichthammol

Cas Number:

8029-68-3

Molecular formula:

unspecified

IUPAC Name:

Ichthammol (UVCB substance)

Test material form:

liquid: viscous

Details on test material:

sample from a batch from regular production released for use as an active pharmaceutical ingredient in medicinal products.

Specific details on test material used for the study:

batch no. R 954623 from regular production at the registrant's manufacturing site.

Ichthammol is characterized in the study report with trademark ICHTHYOL.

Release of substance was based on testing according to pharmacopoeia specifications/instructions of ÖAB / PH.EUR or DAB 10. A certicate of analysis is attached to the original study report.

Sampling and analysis

Analytical monitoring:

not required

Remarks:

No analysis of samples was conducted as ICHTHYOL (Ichthammol) is known to be stable as an aqueous solution.

Details on sampling:

Samples were taken both at the start and the end of the study (after 4 days exposure period) from the tanks of all the concentrations and the control and were stored at LPT (testing facility) at 20°C or colder. However, no analysis of the samples was conducted as ICHTHYOL (Ichthammol) is known to be stable as an aqueous solution.

Test solutions

Vehicle:

no

Details on test solutions:

The test substance was used as supplied. The following concentrations of Ichthyol (Ichthammol) were employed in the exposure medium: 5.0 mg, 8.89 mg, 15.8 mg, 28.1 mg and 50.0 mg Ichthyol/l. The concentrations chosen were based on a preliminary test.

Test organisms

Test organisms (species):

Leuciscus idus

Details on test organisms:

Gold orfe (Leuciscus idus) was chosen for the test; fry of about 6.0 ± 2.0 cm were selected (breeder: FISCHZUCHT EGGERS, D-24594 Hohenwestedt).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

The aim of the present test was to obtain information on the concentration of the test substance in water which kills 50% of the batch of fish (LC5O) within a 24-, 48-, 72- and 96-hour exposure.

Test conditions

Hardness:

Concentration of minerals:
CaCl2 : 2.00 mmol/1
MgSO4 : 0.50 mmol/1
NaHCO3 : 0.77 mmol/1
KCl : 0.077 mmol/l
The sum of calcium and magnesium ions was 2.5 mmol/l, the proportion of Ca:Mg ions was 4:1 and of Na:K ions 10:1.
The acid capacity was about 0.8 mmol/l.
The total hardness was equivalent to 250 mg CaCO3/l.
The total alkalinity was about 0.8 mmol/l.

Test temperature:

20.6°C - 21.2°C (measured daily)

pH:

7.44 - 8.08 (measured daily)

Dissolved oxygen:

not less than 60% of the maximum air saturation value throughout the test (measured daily):
77.8%-93.8%

Salinity:

not applicable

Conductivity:

not measured

Nominal and measured concentrations:

The substance is readily soluble and stable in water. Nominal concentrations, therefore, can be assumed to be in line with actual concentrations.

Details on test conditions:

TEST SYSTEM
The test was carried out as a static test.
The test volume was 10 l per concentration, the concentrations used in this test were 5.0 mg, 8.89 mg, 15.8 mg, 28.1 mg and 50.0 mg Ichthyol/l reconstituted water. 10 fish per tank and concentration were used.
A control tank contained the medium (reconstituted water), also 10 fish were used.

TEST CONDITIONS
Length of fish tested: 6.0 ± 2.0 cm
Number of fish: 10 per concentration and tank
Duration of study: 4 days of exposure
Tank volume: 10 l
static system: water was not changed in 96 hours
Feeding: none, withdrawal 24 hours before start of the test
Light: 16 h photoperiod daily
Aeration: by Pasteur pipettes (supplier: KROLL, D-22525 Hamburg) at a rate of 2 to 3 air bubbles per second
Temperature: 20.6°C - 21.2°C (measured daily)
pH value: 7.44 - 8.08 (measured daily)
Oxygen concentration: not less than 60% of the maximum air saturation value throughout the test (measured daily): 77.8%-93.8%

CONCENTRATION OF MINERALS
CaCL2 : 2.00 mmol/l
MgSO4 : 0.50 mmol/l
NaHCO3 : 0.77 mmol/l
KCl : 0.077 mmol/l
The sum of calcium and magnesium ions was 2.5 mmol/l, the proportion of Ca:Mg ions was 4:1 and of Na:K ions 10:1.
The acid capacity was about 0.8 mmol/l.
The total hardness was equivalent to 250 mg CaCO3/l. The total alkalinity was about 0.8 mmol/l.

EQUIPMENT and control apparatus
Experimental equipment: 10l capacity glass tanks (size: 290 mm (length) x 190 mm (width) x 195 mm (hight)) and normal laboratory equipment
Illumination: by automatic time switch
O2 meter: OXI DIGI530 with oxygen electrode
pH meter: pH 90 with glass electrode (WTW, D-79809 Weilheim)
Temperature stabilised air-conditioning plant: continuous registration
Mercury thermometer: 0.1°C subdivisions, for test water measurement
The pH, temperature and dissolved oxygen concentration of the test water were measured daily.

OBSERVATIONS
Assessment of the behaviour, appearance and mortality of the fish was carried out at the beginning of the study, after 5, 15 and 30 minutes and 1, 3, 6, 24, 48, 72 and 96 hours after the start of the exposure.
Records were kept on any symptoms of toxicological influence (any changes, e.g. different swimming behaviour, different reaction to external stimuli, changes in appearance of the fish) and on death. Dead fish were removed from the tanks as quickly as possible.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

LC50

Remarks:

The nominal LCso (50% mortality of the fish after 96 hours) was calculated by linear regression analysis. The 95% confidence limits were determined according to FINNEY.

Effect conc.:

27.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (total fraction)

Basis for effect:

mortality (fish)

Details on results:

Under the present test conditions no toxic effects caused by Ichthyol (Ichthammol) were observed at a concentration of 5.00 and 8.89 mg Ichthyol/l exposure medium.
First intolerance reactions caused by Ichthyol were observed at a concentration of 15.8 mg Ichthyol/l exposure medium; two animals remained predominantly at the water surface with a marked oblique position. At concentrations from 28.1 mg Ichthyol/l exposure medium onwards the fish remained predominantly at the water surface with a marked oblique position and marked increased gill breathing. The number of affected animals increased in a concentration-related way.
The lowest lethal concentration for fish was 28.1 mg Ichthyol/l exposure medium. The LC50 (96 hrs) was calculated as 27.2 mg Ichthyol/l exposure medium.

Summary of the results
Threshold level of lethal effects: 28.1 mg Ichthyol/l exposure medium
Threshold level of observed effects: 15.8 mg Ichthyol/l exposure medium
No effect concentration (NOEC): 8.89 mg Ichthyol/l exposure medium
The LC50 (96 hours) was calculated as 27.2 mg Ichthyol/l exposure medium

Results with reference substance (positive control):

not applicable

Reported statistics and error estimates:

The nominal LC50 (50% mortality of the fish after 96 hours) was calculated by linear regression analysis. The 95% confidence limits were determined according to FINNEY.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Test system: Fish, acute toxicity, static test
EC guideline: L383A: C.l.
OECD guideline: 203
Test substance: Ichthyol (trademark name for Ichthammol)
Species: Gold orfe/Leuciscus idus
Test period: 96 hours
Dose level: 5.00 mg/l, 8.89 mg/l, 15.8 mg/l, 28.1 mg/l and 50 mg/l
Threshold level of lethal effects: 28.1 mg IchthyoI/l exposure medium
Threshold level of observed effects: 15.8 mg Ichthyol/l exposure medium
No effect concentration (NOEC): 8.89 mg Ichthyol/l exposure medium
The LC50 (96 hours) was calculated as 27.2 mg Ichthyol/I exposure medium