1-Propanaminium, 3-amino-N-(carboxymethyl)-N,N-dimethyl-, N-C8-10 (even numbered) acyl derivs., hydroxides, inner salts

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:

2014-11-03 to 2014-11-10

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

The limit concentration and the control were analytically verified from freshly prepared test media after 0 and 72 h and from corresponding 24 h aged test media after 24 and 96 h.
Analytical evaluation of the test item was carried out via LC-MS/MS.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
The stock solution was prepared by directly weighing the test item into the test medium. The test media were mixed with an ultraturrax (1 min, 17000 rpm).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Strain: Danio rerio
- Source: Umweltbundesamt, Schichauweg 58, 12307 Berlin, Germany
- Length at study initiation (mean): 2.5 cm
- Weight at study initiation (mean): 0.227 g
- Feeding during test: no; the test fish were not fed 24 h before test start

ACCLIMATION
- Acclimation conditions (same as test or not): Tap water; 23 ± 2°C, diffuse light (0.1 - 10 µmoI . m-2 . s-1, natural photoperiod); water changes at least once per week; dissolved oxygen concentration > 80 % of the air saturation value
- Type and amount of food: Sera Vipan; SERA GMBH, 52518 Heinsberg, Germany; the amount of food was 4% of the fish body weight per feeding day
- Feeding frequency: 3 times per week
- acclimation: at least 12 days; fish with mortality < 5 % within these days prior to start of the exposure were used in the test

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

231 mg/L as CaCO3

Test temperature:

22.9 ± 0.3

pH:

7.46 - 7.93

Dissolved oxygen:

80 - 100%

Nominal and measured concentrations:

nominal: 0, 100 mg a.i./L
measured concentrations were in the range of 99-108%

Details on test conditions:

TEST SYSTEM
- Test vessel: 3 L glass aquaria, filled with 2 L of natural river water (dimensions: 11.5/13/20 cm, depth of water: at least 13.5 cm), and covered with glass plates
- Aeration: yes
- Renewal rate of test solution (frequency): daily
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.79 g fish per litre test solution

TEST MEDIUM / WATER PARAMETERS
river water as specified in section "any other information on materials and methods"
The natural river water was stored at 15 ± 2°C. At least 24 h before use, the natural river water was temperated to 23 ± 2°C and aerated.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 9.5 hours light
- Light intensity: 0.42 to 0.64 (mean value 0.52) µmol photons : m-2 . S-1

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
mortality at 24, 48, 72 and 96 h
Records were kept of visible abnormalities (e.g. loss of equilibrium, swimming behaviour, respiratory function, pigmentation, etc.)

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: The limit concentration was extrapolated from the results of a preliminary range finding test conducted under semi-static conditions, with natural river water and the nominal Betaine concentrations of 34.6 and 3.46 mg a.i./L. As no mortality was observed in the limit concentration, no additional testing of further concentrations was carried out.

Reference substance (positive control):

not required

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Details on results:

- Mortality of control: 0/7

Results with reference substance (positive control):

n.a.

Validity criteria fulfilled:

yes

Conclusions:

The 96 h LC50 of C8-10 Alkylamidopropyl betaine was > 100 mg a.i./L in zebrafish (Danio rerio) using natural river water.

Executive summary:

In a 96 h acute toxicity study according to OECD guideline 203, adopted 17 July 1992, zebrafish (Danio rerio) were exposed to C8-10

Alkylamidopropyl betaine at nominal concentrations of 0 (control) and 100 mg/L under semi-static conditions in natural river water containing 3.46 mg/L TOC (2.71 mg/L DOC). Measured test item concentrations were in agreement with nominal (99-108%). These concentrations remained stable during both renewal periods. Given these results, effect parameters can be based on the nominal test concentrations.

No mortality or other clinical effect was observed at the limit concentration and the control during the 96-hour test period. The 96 h LC50 was >100 mg a.i./L. 

This study is classified as acceptable and satisfies the guideline requirements for an acute toxicity study with Danio rerio..

 

Results Synopsis

Test organism size/age (mean wet weight or length): Zebrafish (Danio rerio) (2.5 cm / 0.227 g)

Test Type: Static Renewal

LC50:  >100 mg a.i./L

Endpoint(s) Effected: mortality

Description of key information

96 h LC50 >100 mg a.i./L (analytically verified nominal concentration); OECD TG 203, Danio rerio, semi-static, GLP, RL1; natural river water containing 3.46 mg/L TOC

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

In a 96 h acute toxicity study according to OECD guideline 203, adopted 17 July 1992, zebrafish (Danio rerio) were exposed to C8-10 Alkylamidopropyl betaine at nominal concentrations of 0 (control) and 100 mg/L under semi-static conditions in natural river water containing 3.46 mg/L TOC (2.71 mg/L DOC). Measured test item concentrations were in agreement with nominal (99-108%). These concentrations remained stable during both renewal periods. Given these results, effect parameters can be based on the nominal test concentrations.

No mortality or other clinical effect was observed at the limit concentration and the control during the 96-hour test period. The 96 h LC50 was >100 mg a.i./L. 

The following results were obtained with the closely related read-across substances C8-18 and C18 unsatd. AAPB, C8-18 AAPB and Formamidopropylbetaine (included in the dossier to support the read-across for other endpoints):

The acute toxicity of C8-18 and C18 unsatd. AAPB to fish (Pimephales promelas) was investigated in a study conducted according to OECD Guideline 203 and EU Method C.1. A 96 h LC50=1.11 mg a.i./L nominal (=3.7 mg product/L) was determined.

In a 28-d toxicity study conducted according to OECD Guideline 204 under flow-through conditions and using rainbow trouts (Oncorhynchus mykiss) as test organism, the 28-d NOEC and LOEC of C8-18 AAPB were 0.16 and 0.5 mg a.i./L nominal based on mortality and sublethal effects.

Formamidopropylbetaine induced no visible effects in carp at 100 mg/L (NOEC). The 96h-LC50 was> 100 mg/L based on analytically confirmed nominal concentrations.