Reaction mass of 2-O-α-L-Rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoic acid and α-L-Rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoic acid

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:

2016-03-14 - 2018-03-18 (experimental phase)

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

Organisation for Economic Cooperation and Development. OECD Guidelines for the Testing of Chemicals, "Fish Acute Toxicity Test", Procedure 203, adopted 17 July, 1992.

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Version / remarks:

Commission Regulation (EC) No 440/2008 of 30 May 2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH), C.1. Acute Toxicity for Fish (REACH) (O.J. L 142 of 31.5.2008).

Deviations:

no

GLP compliance:

no

Remarks:

, but the study was conducted in accordance with ISO/IEC 17025.

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of 2-O-α-L-Rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoic acid and α-L-Rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoic acid

EC Number:

604-610-3

Cas Number:

147858-26-2

IUPAC Name:

Reaction mass of 2-O-α-L-Rhamnopyranosyl-α-L-rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoic acid and α-L-Rhamnopyranosyl-β-hydroxydecanoyl-β-hydroxydecanoic acid

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No. of test material: Sponsor

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature, protected from light
- Stability under test conditions: Stable under storage conditions

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Remarks:

direct weighing

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Since the test item is soluble, the test concentration was prepared by direct addition of the test item to natural water. This mixture was moderately stirred for 5 minutes. The resulting solution was used as the test medium (one test vessel).
- Controls: A blank consisting of natural water only, without any addition of test item or other additives, served as a control (one test vessel).
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): none stated.

Test organisms

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Common name: Guppy
- Source: The fish were born and bred in the aquaria of the laboratories of the Arcadis Schweiz LTD in Schlieren.
- Age at study initiation (mean and range, SD): young
- Length at study initiation (length definition, mean, range and SD): target: length of 1–3 cm
- Weight at study initiation (mean and range, SD): not stated, (target: total loading ≤ 1 g fish weight/l)
- Method of breeding:
- Maintenance of the brood fish:

ACCLIMATION
- Acclimation period: Young fish are held under quarantine conditions in 40 l glass aquaria until use in the test, duration: 7 days prior to start of exposure
- Acclimation conditions (same as test or not): yes
- Health/mortality: No mortality was observed during the 7 day period prior to the start of the exposure (requirement: total mortality <5% during the 7 day period prior to the start of the exposure –otherwise the fish cannot be used).

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

none

Test conditions

Hardness:

Parameter Unit Minimum Maximum
Total Hardness °f H 20.6 24.2
mmol/l 2.06 2.42
mg/l 206 242
CO3 Hardness °f H 18.6 21.7
mg/l 186 217
Residual Hardness °f H 2.0 3.0
mg/l 20 30

Test temperature:

Target: 21–25 °C, controlled by a thermostat at ±2°C

pH:

7.4 - 7.8 (water)
pH was adjusted to pH 6.5–7.0.

Dissolved oxygen:

The total dissolved oxygen content is kept at >60% of the saturation value with aeration via Pasteur pipettes

Salinity:

Nitrate mg/l 7.3
Chloride mg/l 16.1
Sulfate mg/l 18.6

Conductivity:

477 - 536 μS/cm (room temperature)

Nominal and measured concentrations:

nominal: 100 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass aquaria filled with 1.0 liter of water
- Aeration: The total dissolved oxygen content is kept at >60% of the saturation value with aeration via Pasteur pipettes
- Renewal rate of test solution (frequency/flow rate): none
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: total loading ≤ 1 g fish weight/l

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Continuously aerated natural water of the town of Schlieren
- Chlorine: 16.1 mg/l
- Culture medium different from test medium: no
- Intervals of water quality measurement: dependent on parameter, bimonthly, monthly or yearly

OTHER TEST CONDITIONS
- Adjustment of pH: yes
- Photoperiod: 16 h a day
- Light intensity: 16 h photoperiod a day, supplied by overhead white fluorescent tubes, no transition period

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Oxygen, pH, temperature: Measurement were made prior to the addition of the fish to the test solutions and after 2, 24, 48, 72 and 96 h of exposure.
Observations: Observations of any potential abnormal behaviour of the fish were made after 2, 24, 48, 72 and 96 h of exposure. Mortality of the fish was recorded at the same time intervals. Any dead fish were removed immediately from the test media, and their respective weight and length were recorded.
Fish length and weight: At the end of the test, length and weight of all surviving fish were also recorded.

TEST CONCENTRATIONS
- Results used to determine the conditions for the definitive study: The nominal test concentration of 100 mg/l was chosen based on preceding tests with Algae (OECD 201) and Daphnia (OECD 202) in which EC50 values of >100 mg/l were observed

Reference substance (positive control):

not required

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Key result

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: The one fish that died is not considered to be a relevant effect as defined by the OECD guidance.

Effect concentrations 3

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: The one fish that died is not considered to be a relevant effect as defined by the OECD guidance.

Details on results:

- Behavioural abnormalities: none
- Observations on body length and weight: none differing from control
- Other biological observations: none differing from control
- Mortality of control: none
- Other adverse effects control: none
- Abnormal responses: none
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: none
- Effect concentrations exceeding solubility of substance in test medium: none

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The study was conducted according to OECD guideline 203 and EU method C.1 on the registered substance itself. The method is to be considered scientifically reasonable with no deficiencies in documentation or deviations from the guidelines, the validity criteria were met. Hence, the results can be considered as reliable to assess the toxicity of test item towards fish. At the nominal concentration of 100 mg/l one fish out of a total of seven died after 96 h of exposure. No fish out of seven died in the blank control. The no-observed-effect concentration (NOEC, LC0) is 100 mg/l nominal concentration since the one fish that died is not considered to be a relevant effect as defined by the OECD guidance. The LC50 of the test item to Poecilia reticulata is hence >100 mg/l nominal concentration.
The criteria for classification of a substance in category Acute 1 are defined on the basis of acute aquatic toxicity data only (EC50 or LC50); classification is required for values ≤ 1 mg/L. Therefore, the substance does not need to be classified for acute (short-term) aquatic hazard.
Further, classification as aquatic chronic category 3 or worse is required if the 96 hr LC 50 (for fish), and/or 48 hr EC 50 (for crustacea) and/or 72 or 96 hr ErC 50 (for algae or other aquatic plants) values are > 100 mg/l (and the substance is not rapidly degradable and/or the experimentally determined BCF ≥ 500 (or, if absent, the log Kow ≥ 4).
The determined 96h LC50 is > 100 mg/l, so no classification as aquatic chronic Cat. 3 or worse is required.

Executive summary:

The acute lethal toxicity of the test item to Poecilia reticulata (Guppy) was investigated in a limit test under static exposure conditions over a period of 96 h in an OECD 203 study.

Since the test item is soluble, the test solution was prepared by direct addition of the test item to natural water.

The only applied nominal concentration of the test item was 100 mg/l. Therefore, this test represents a limit test.

Seven fish were exposed to the test item, and 7 fish were used in the blank control.

No analytical verification of the test item was conducted. The evaluation of the test was based on the nominal concentration of the test item, assuming the test item to be stable in water over 96 h.

At the nominal concentration of 100 mg/l, one fish out of a total of seven died after 96 h of exposure. No fish out of seven died in the blank control.

Toxic effects other than mortality, e.g. loss of coordination, hypo- or hyperactivity and swimming on the back, were not observed.

The no-observed-effect concentration (NOEC, LC0) is 100 mg/l nominal concentration since the one fish that died is not considered to be a relevant effect as defined by the OECD guidance.

The LC50 of the test item to Poecilia reticulata is >100 mg/l nominal concentration.

100% mortality (LC100) could not be detected.

The substance does not need to be classified according to Regulation 1272/2008 and amendments.