Amides, from C8-10-fatty acids and tetraethylenepentamine

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

July 5 2007 to May 5 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: OECD 203 guideline study, GLP, All validity criteria fulfilled, complete identification of test substance, including chemical analyses

Justification for type of information:

Refer to read across justification document in section 13

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures. OECD Environmental Health and Safety Publication, Series on Safety testing and assessment, No. 23, Paris (December 2000)

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Storage of samples: none
Samples treatment: dilution in methanol with a dilution factor ranging from 2 to 10

Vehicle:

no

Details on test solutions:

Osmotically filtered water was used to prepare the dilution water. Chemical reagents used for the preparation of dilution water were of analytical grade. For the preliminary test, solutions of IMIDAZOLINE 4900 in the dilution waster were prepared 72h before the test was started. For the definitive test, stock solutions of IMIDAZOLINE 4900 were prepared in the dilution waster by stirring during 72 h for the first day solutions and 24 h for the three others. Solutions were filtered through HAWP0047 millipore membranes to obtain limpid solutions before the test was started.

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The organisms used for thes test were Danio rerio (Teleostei, Cyprinidae). The batch no 07/Br/01/1 was supplied by HB development (La Fond Garei, 69490 Saint-Forgeux, France). The fish were placed under the test conditions (medium, identical or lower biological loading, temperature, lighting, etc.) 20 days before the beginning of the test. Fish were fed with TetraPro (Tetra GmbH, D-49304 Melle, Germany). The fish selected for the test were all from the same batch and were of the same age (date of birth stated by the supplier: 20/02/07). Their size (determined by measuring 10 randomly sampled fish) was 3.1 +- 0.1 cm. The mortality in the aquarium containing the batch of fish used for the tests was 5 % over the 7 days preceeding the test.
The sensitivity of the biological reagent is checked at least once for each new batch of fish, by determining the 24h LC50 of the potassium dichromate. For reference, the result of the last test, carried out on 19/07/07, was 24h LC50 of 270 mg/L (potassium dichromate Aldrich batch no 18703HB).

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Remarks on exposure duration:

with renewal of water every 24 hours

Hardness:

Sum of Ca and Mg ions in dilution/rearing waster is 2.5 mM and ratios of Ca/Mg and Na/K are 4 and 10 respectively

Test temperature:

21.5 +- 0.5 deg celsius

pH:

7.49 - 8.52 (preliminary test)
7.70 - 8.09 (definitive test)

Dissolved oxygen:

76 - 98 % (preliminary test)
86 - 98 % (definitive test)

Salinity:

not measured - freshwater test

Nominal and measured concentrations:

nominal: 0.46; 0.67; 0.96; 1.39; 2.00 mg/L
measured: 0.09; 0.20; 0.33; 0.39; 0.64 mg/L

Details on test conditions:

The study was carried out in a themostatically controlled room. The temperature was measured on a a continuous basis in a test flask. The room was equipped with a programmed lighting system which produced a controlled lighting sequence: intensity between 400 and 800 lux at the surface of the aquaria; photo period of 12 hours of darkness per 24 h.

Physico-chemical parameters were measured using a METTLER TOLEDO 345 pH meter for measurement of pH and with a WTW OXI 538 oxymeter for dissolved oxygen measurements.

The study was performed using aquariums with a nominal capacity of 5 liters, covered with Altuglas cover plates drilled with holes to allow the test solutions to be aerated. Volume of test medium per aquarium was 2.5L

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

0.31 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

0.21 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

72 h

Dose descriptor:

LC50

Effect conc.:

0.2 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

0.19 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

The appearance of the test solutions was visually checked at the beginning and at the end of the test. Solutions were found to be clear, colourless over the period of the test. No precipitation was observed at the end of the test.

For the calculation of the effective concentrations, the concentrations were measured before and after renewal of test solution, and then a geometric mean was calculated, for each of the duplo test concentrations. An arithmetic mean was then calculated from the duplos to get the effective concentration.

There were no behavioural abnormalities reported in this study

Results with reference substance (positive control):

24 hLC50 of 270 mg/L

Reported statistics and error estimates:

The lethality data were analysed using an Excel sheet program. It was designed to calculate the LC50 values using Probita analysis and the 95% confidence interval.

Sublethal observations / clinical signs:

The percentage of dead fish were counted everyday for preliminary and definitive tests, and are shown below

Preliminary test: percentage of dead fish

Nominal concentrations (mg/L)	Mortality (%)
	24 h	48 h	72 h	96 h
50	100	100	100	100
10	100	100	100	100
5	100	100	100	100
1	20	40	90	80
0.5	0	0	0	0
0.1	0	0	0	0
0	0	0	0	0

Definitive test: percentage of dead fish

Extrapolated concentrations (mg/L)	Mortality (%)
	24 h	48 h	72 h	96 h
0.64	100	100	100	100
0.39	100	100	100	100
0.33	20	100	100	100
0.20	0	10	20	30
0.09	0	0	0	0
0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of IMIDAZOLINE 4900 to D. rerio was assessed under semi-static conditions according to the OECD 203 guideline, and resulted in the following LC50 values
24 h LC50 = 0.31 mg/L; 48 h LC50 = 0.21 mg/L; 72 h LC50 = 0.20 mg/L; 96 h LC50 = 0.19 mg/L

Executive summary:

Summary

The acute toxicity (assessed by lethality) of the test item IMIDAZOLINE 4900 to Danio rerio (freshwater fish) for a duration of 96 hours was assessed according to the OECD guideline 203.

Fish were exposed to a range of concentrations of IMIDAZOLINE 4900 dissolved in dilution water. The definitive test was performed under semi-static conditions, with a renewal of test solutions every day. The measured toxic effect was the mortality after 24, 48, 72, and 96 hours.

The effective concentrations killing 50% of fish at 24h, 48, 72h, and 96h (LC50 -24h, 48h, 72h and 96h) were calculated by regression analysis using the Probit/log model. The results are presented in the following table.

Duration of exposure period (h)	LC50 value, mg/L (95% CI)
24	0.31
48	0.21
72	0.20
96	0.19 (ND)

ND: not determined

The study was performed in compliance with the folllowing quality criteria:

• The mortality in the control did not exceed 10% at the end of the test
• The concentration of dissolved oxygen in the test vessels remained above 60% of the air saturation value at the end of the test
• The pH did not vary by more than one unit.

Description of key information

Read-across from 68442 -97 -7 as structural analogue.

Based on the limited number of acute fish data it can be observed that fish are not more sensitive than daphnia and algae to amidoamines/imidazolines.

For the calculation of the PNECaquatic bulk it is considered unlikely that fish toxicity will be critical for amidoamines/imidazolines category. Hence any additional toxicity testing with fish will not add scientific value to the ecotoxicity profile of this category other than for obtaining a lower assessment factor. It is therefore concluded that for scientific reasons and in accordance to REACH legislation further testing on fish has to be avoided for reasons of animal welfare and that based on the weight of evidence available on ecotoxicity data for several cationic surfactant types the use of a safety factor of 10 is considered justified for the derivation of the PNECaquatic,bulk.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.19 mg/L

Additional information

The limited number of short-term toxicity fish tests available were conducted in a period when no reliable specific method of analyses was available. The concentrations were therefore in old studies not analytically verified. The reliability of these results is limited because of the poor solubility of the test substances and partial sorption onto the walls of test vessels. In one study however the exposure concentration was quantified and this study has been selected a Key study. The other two were considered to be valid with restrictions and can be used as supporing studies.