Reaction products of 1,4-Benzenedimethanol and 1-naphthol

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:

06 May 2013 to 10 May 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Ministry of Economy Trade and Industry (METI),

Version / remarks:

Guidelines for studies on the new chemical substance as required by the Law Concerning the Evaluation of Chemical Substances and Regulation of their Manufacture, etc (Chemical Substance Control Law) 1973, amended 2009 under the reference of YAKUSHOKHATSU No. 1121002, SEIKYOKU No.2 and KANPOKIHATSU No. 021121002 and partially amended 2006 as the joint ordinance of The Japanese Ministry of Economy Trade and Industry (METI), Ministry of Health, Labour and Welfare (MHLW) and Ministry of the Environment (MOE).

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: JMAFF 2-7-1-1

Version / remarks:

The guidelines related to the study reports for the registration application of pesticide (Ref. No. 12-Nousan-8147 on 24 November 2000) & Ref. No.13-Seisan-3986 on 10 October 2001.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

No further details specified in the study report

Analytical monitoring:

yes

Details on sampling:

Analytical measurement was performed at the applied test concentration level on day 0, 1, 3 and 4, corresponding to the first and last renewal period during the test and at the control at the beginning and at the end of the test. At each occasion, three replicate samples were taken from the test solutions and one sample from the control solution.
The samples were analysed by HPLC-UV method.

Vehicle:

no

Details on test solutions:

Because the test item is very poorly soluble in water, a test solution was prepared using a saturated solution method. A supersaturated test item stock solution (nominally 100 mg/L) was prepared by dispersing/dissolving the amount of test item into the test medium (aquarium water) two days before the start of the study. This solution was shaken for about 24 hours at approximately 30°C and then was equilibrated for about 24 hours at the test temperature. The non-dissolved test material was removed by filtration through a fine (0.22 μm) filter to give the 100 % v/v saturated solution.
As limit test was carried out, further dilution of stock solution was not performed.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

Species: Zebra-fish (Brachydanio rerio)
Source: Ágnes Németh (self-employed), Fish Farm, Veszprém, Hungary
Justification of species: Zebra-fish (Brachydanio rerio) is one of the convenient species for acute fish toxicity test.
Number of animals: seven fish per test group
Body length of animals: 2.6 – 3.1 cm
Food and Feeding: The fish were not fed during the test
Acclimatisation: >12 days
Animal health: 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. There was no mortality of the population for seven days before the test.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Not specified

Hardness:

The hardness of the dilution water was determined as 54 mg/L (as CaCO3).

Test temperature:

The test temperature was between 20.8 and 21.2 °C.

pH:

The pH was in the range of 8.02 – 8.20.

Dissolved oxygen:

The dissolved oxygen concentration was in the range of 78 – 98 % of the air saturation value at the temperature used

Salinity:

Not specified

Conductivity:

Not specified

Nominal and measured concentrations:

Because no toxic effect was observed in the preliminary range-finding test at saturation, only this concentration (100 % v/v saturated solution) and one control were tested in a limit-test.

Details on test conditions:

The test duration was 96 hours. One aquarium was used for test group and control group respectively in a semi-static system. Each aquarium comprised 7 fish and 5 L test solution. The animals were not fed during the test. The loading of the test aquaria was less than 1.0 g fish/L test solution at the start of the experiment.
The choice of the test concentrations was done on the basis of the results of the preliminary range-finding test.

Reference substance (positive control):

no

Key result

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LOEC

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Key result

Duration:

96 h

Dose descriptor:

LC100

Effect conc.:

> 100 other: % v/v saturated solution

Nominal / measured:

meas. (initial)

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mortality (fish)

Details on results:

The achievable validity criteria were within acceptable limits and therefore the study can be considered as valid.

Results with reference substance (positive control):

Not application - positive control was not used.

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects. The LC50, NOEC, LOEC and LC100 were determined directly from the raw data.

Sublethal observations / clinical signs:

ANIMAL WELFARE

After the experiment, all fish were humanely killed. Euthanasia for all animals was applied by an overdose of an aqueous solution of 250 mg/L Tricaine Methanesulphonate (MS 222; CAS No.: 886-86-2). This technique is considered to provide a humane euthanasia by direct depression of the brain and vital centers.

MORTALITY DATA

Cumulative mortality data from the exposure of fish to the test material during the definitive test are given below:

Cumulative mortality data in the definitive test

Test group	Mortality rate of treated fish (Dead fish/Treated fish)
	3h	6h	24h	48h	72h	96h
Control	0/7	0/7	0/7	0/7	0/7	0/7
100% v/v saturated solution	0/7	0/7	0/7	0/7	0/7	0/7

BODY WEIGHT

The body weight of 7 fish was weighed at the start of the test. The measured and calculated data are listed below:

Measured and calculated data of bodyweight

Test Group	Measured weight of 7 fish (g)	Calculated mean weight of 1 fish (g)	Loading of testing aquarium (g fish/L testing liquid)
Control	2.17	0.31	0.43
100% v/v saturated solution	2.05	0.29	0.41

There was no considerable difference observed concerning body weights between the groups.

Validity criteria fulfilled:

yes

Conclusions:

The test item SN-475N had no toxic effect at aquatic saturation on fish; the LC50 results and the LOEC are higher than the solubility level of the test item in the test medium.

Executive summary:

The acute toxicity of SN-475N was assessed with acute fish toxicity test on Zebra-fish (Brachydanio rerio), over an exposure period of 96 hours in a semi-static system.

Toxic effects were not observed during the preliminary concentration range-finding test therefore a limit test was performed in the main experiment using only one test concentration at the limit of solubility of the test item in the test medium (100 % v/v saturated solution) and one control group.

Test concentration was analytically determined on day 0, 1, 3 and 4, corresponding to the first and last renewal period, however the measured concentration was below the Limit of Quantification (LOQ = 2.0 mg/L) also at the start and at the end of the test, after increasing the concentration by a factor of 25 by freeze-drying method. Therefore it can be stated that the solubility level of the test item is lower than 2.0/25=0.08 mg/L.

All achievable validity criteria were met during this study.

Under the conditions of this acute fish toxicity study on Zebra-fish (Brachydanio rerio) the observed endpoints for the effect of SN-475N were the followings:

The 24h, 48h, 72h and 96h LC50 value: > 100 % v/v saturated solution

The 96h NOEC: 100 % v/v saturated solution

The 96h LOEC: > 100 % v/v saturated solution

The 96h LC100 value: > 100 % v/v saturated solution

Based on the results of this study, the test item SN-475N had no toxic effect at aquatic saturation on fish; the LC50 results and the LOEC are higher than the solubility level of the test item in the test medium.

Description of key information

The test item SN-475N had no toxic effect at aquatic saturation on fish; the LC50 results and the LOEC are higher than the solubility level of the test item in the test medium, > 100% v/v saturated solution.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L