Fatty acids, montan-wax, stearyl esters

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

04 - 08 Feb 2013

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

17 Jul 1992

GLP compliance:

yes (incl. QA statement)

Remarks:

GYEMSZI, National Institute for Quality and Organizational Development in HealthCare and Medicine, Hungary (08 - 11 Oct 2012)

Analytical monitoring:

no

Remarks:

The test item concentrations could not be determined due to very low solubility in water (< LOQ).

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: A saturated solution method was used to prepare test solutions because the test item is very poorly soluble in water. A supersaturated test item stock solution of 100 mg/L (nominal) was prepared by dispersing/dissolving the required test item in test medium (aquarium water) 2 d before the start of the study. This solution was shaken for 24 h at approximately 30 °C and then was equilibrated for about 24 h at test temperature. The non-dissolved test material was removed by filtration through a fine filter (0.22 µm) to give the 100% v/v saturated solution.
- Controls: Dilution water without test item (circulated, filtered and cooled).

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: Rainbow trout
- Source: Hoitsy & Rieger Kft., Trout Farm, Miskolc-Lillafüred, Hungary
- Length at study initiation: 6.0 - 6.5 cm
- Calculated mean weight of 1 fish at study initiation: 2.79 g (control), 2.67 g (treatment). There was no considerable difference in body weights between the groups.
- Method of breeding: Fish were bred in a well-known fish farm, under disease- and parasite-controlled conditions. Fish were observed during the acclimatisation and test periods, the fish were healthy.
- Maintenance of the brood fish: Mortalities were < 5% of the population 7 d before the test.

ACCLIMATION
- Acclimation period: > 12 d

FEEDING DURING TEST
- Frequency: The fish were not fed during the test.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

203.5 mg/L as CaCO3

Test temperature:

13.7 - 14.5 °C

pH:

7.32 - 8.01

Dissolved oxygen:

94 - 98% of the air saturation value

Nominal and measured concentrations:

100% v/v saturated solution (nominal: 100 mg/L)

Details on test conditions:

TEST SYSTEM
- Test vessel: Aquaria filled with 20 L test solution
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: < 1.0 g fish/L

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement: Daily (pH, temperature, oxygen saturation)

OTHER TEST CONDITIONS
- Photoperiod: 16 h light, 8 h dark

EFFECT PARAMETERS MEASURED:
- Mortality and sub-lethal effects: after 3, 6, 24, 48, 72 and 96 h
- Body weight: Before test start

TEST CONCENTRATIONS
- Range finding study : Yes
- Test concentrations: 0.1, 1, 10, and 100% v/v saturated solution
- Results used to determine the conditions for the definitive study: Yes, the test concentrations were selected on the basis of the results of the preliminary range-finding test: 0% mortality at 100% v/v saturated solution after 96 h.

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

100% v/v saturated solution

Duration:

96 h

Dose descriptor:

NOELR

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

100% v/v saturated solution

Duration:

96 h

Dose descriptor:

LOELR

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other:

Remarks:

100% v/v saturated solution

Details on results:

- Observations on body length at the end of the test: 6.0 - 6.4 cm (control), 6.1 - 6.5 cm (treatment)
- Mortality of control: 0%
- Effect concentrations exceeding solubility of substance in test medium: No

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects.
The LL50, NOELR, LOELR and LL100 were determined directly from the raw data.

Sublethal observations / clinical signs:

VALIDITY CRITERIA

The validity criteria were within the acceptable limits (Table 1). Therefore, the study is valid.

 

Table 1: Validity criteria for OECD 203.

Guideline criterion	Outcome	Validity criterion fulfilled
The mortality in the control(s) should not exceed 10% (or one fish if less than ten are used) at the end of the test	Mortality in the control after 96 h: 0%	Yes
The dissolved oxygen concentration must have been at least 60 per cent of the air saturation value throughout the test	Dissolved oxygen concentrations were 94 – 98% of the air saturation value during the test.	Yes
There must be evidence that the concentration of the substance being tested has been satisfactorily maintained, and preferably it should be at least 80 per cent of the nominal concentration throughout the test. If the deviation from the nominal concentration is greater than 20 per cent, results should be based on the measured concentration.	According to the obtained results of the validation, the test item has very low solubility in water below the Limit of Quantification. Therefore, the test item concentrations could not be determined.	not applicable

 

BIOLOGICAL RESULTS

The cumulative mortality during the definitive test is given in Table 2.

 

Table 2. Mortality data.

test group	mortality rate of treated fish (dead fish/treated fish)
	3 h	6 h	24 h	48 h	72 h	96 h
control	0/7	0/7	0/7	0/7	0/7	0/7
100 mg/L	0/7	0/7	0/7	0/7	0/7	0/7

 

CONCLUSION

Based on the result, the test item had no toxic effect on fish at aquatic saturation. The LL50 values and the LOELR are higher than the solubility level of the test item in the test medium.

Validity criteria fulfilled:

yes

Remarks:

For further details please refer to “Any other information on results incl. tables”.

Description of key information

No effects up to the limit of water solubility (OECD 203, O. mykiss)

Additional information

There is one experimental study available, in which the toxicity of Fatty acids, montan-wax, stearyl esters (CAS 68308-30-5) to fish was investigated according to OECD guideline 203 and in compliance with GLP.

In a static limit test, the Rainbow trout Oncorhynchus mykiss was exposed to a 100% v/v saturated solution for 96 h. Since the test item is very poorly soluble in water, a supersaturated test item solution was prepared by dispersing a nominal concentration of 100 mg/L test item in test medium 2 d prior test start, shaking for 24 h (at approximately 30 °C) and finally filtering the solution through a 0.22 µm filter prior use. A chemical analysis of test item concentrations in the test solution was not possible due to the very low solubility of the test item in water (below the limit of quantification).

After 96 h, no mortality was observed. Based on the obtained results, Fatty acids, montan-wax, stearyl esters (CAS 68308-30-5) had no toxic effect on fish at aquatic saturation and the derived LL50 (96 h) was > 100 mg/L (above the solubility level of the test item in the test medium).