Methyl DL-serinate hydrochloride

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

09-sep-2014 - 11-sep-2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study has been performed according to OECD and/or EC guidelines and according to GLP principles

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Methyl DL-serinate hydrochloride

EC Number:

227-047-1

EC Name:

Methyl DL-serinate hydrochloride

Cas Number:

5619-04-5

Molecular formula:

C4H9NO3.ClH

IUPAC Name:

methyl 2-amino-3-hydroxypropanoate hydrochloride

Test material form:

solid: crystalline

Details on test material:

Identification : DL-serine Methylester HCl
Molecular formula: C4H9NO3·HCl
Molecular weight: 155.58
CAS Number: 5619-04-5
Roche Number : RO0135435-001
Description: White crystalline powder
Batch: F140001
Purity/Composition: See Certificate of Analysis
Test substance storage: At room temperature in the dark
Stable under storage conditions until: 14 September 2014 (expiry date)
Purity/composition correction factor required: No
General information: Avoid temperature above 60°C
Hygroscopic: Not indicated
Volatile: Not indicated
Stability in water: Not indicated
Solubility in water: >588 g/L (20°C)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples for possible analysis were taken from all test concentrations and the control according to the schedule below.

Frequency at t=0 h and t=48 h
Volume 1.0 ml from the approximate centre of the test vessels
Storage Samples were stored in a freezer until analysis.

At the end of the exposure period, the replicates were pooled at each concentration before sampling.

Additionally, reserve samples of 1.0 ml were taken for possible analysis. If not used, these samples were stored in a freezer for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.

Test solutions

Vehicle:

no

Details on test solutions:

The batch of DL-serine Methylester HCl tested was a white crystalline powder that was completely soluble in test medium at the concentrations tested. No correction was made for the purity/composition of the test substance.

Preparation of test solutions started with a nominal concentration of 100 mg/l applying a short period of vigorous shaking to accelerate the dissolving of the test substance in the test medium. The lower test concentrations were prepared by subsequent dilutions of the 100 mg/l concentration in test medium. The final test solutions were all clear and colourless.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (Crustacea, Cladocera) (Straus, 1820)
- Source: In-house laboratory culture with a known history.
- Age at study initiation: < 24 hours
- Method of breeding: With newborn daphnids, i.e. less than 3 days old, by placing them individually in 50 ml M7-medium.
- Feeding during test: No

BREEDING:
Method: Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by acyclical parthenogenesis under specified breeding conditions. Each batch started with newborn daphnids (i.e. less than 3 days old) and the maximum age of the culture was 4 weeks
Medium: M7-medium.(Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).
Type of food: Supension of fresh water algae.
Feeding frequency: Daily.
Validity of the batch:Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

180 mg/l expressed as CaCO3

Test temperature:

Start of the test:between 20.2 and 20.7°C
During the test: between 19.1 and 19.8°C

pH:

Between 6.8 and 8.1

Dissolved oxygen:

Between 9.2 and 9.4 mg/l

Nominal and measured concentrations:

Nominal concentrations: Control, 0.1, 1.0, 10 and 100 mg/l .
Measured concentrations: Analysis of the samples taken from the highest test concentration at the start and the end of the test showed that the measured concentration was in agreement with nominal (81-111%). Based on this result, the EC50 values were based on the nominal concentration of 100 mg/l.

Details on test conditions:

TESTSYSTEM:
Test vessel: all-glass with 100 ml capicity containing 80 ml of test solution.
Aeration: none
No. of organisms per vessel: 5
No. of vessels per concentration (replicates): 0.1, 1.0 and 10 mg/l 2 replicates and 100 mg/l 4 replicates
No. of replicates per control: 4
Loading: 5 daphnids per 80 ml of test solution

TESTMEDIUM/WATER PARAMETERS
- Source/preparation of dilution water: Adjusted ISO medium: the following chemicals (analytical grade) are dissolved in tap water purified by Reverse Osmosis (RO-water, GEON Waterbehandeling, Berkel-Enschot, The Netherlands):
Macro salts:
CaCl2.2H2O : 211.5 mg/l
MgSO4.7H2O: 88.8 mg/l
NaHCO3 : 46.7 mg/l
KCl : 4.2 mg/l
The hardness: 180 mg/l expressed as CaCO3 and the pH: 7.7 ± 0.3 after aeration.
- Culture medium different from test medium: yes, for culturing M7 medium is used
-Interval of water quality measurements: pH and dissolved oxygen was measured at the beginning of the test and at the end for the control and the highest concentration. Temperature was measured continuously in a temperature control vessel.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : EC50 t=24, 48 h

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 10
- Test concentrations: control and 0.1, 1.0, 10 and 100 mg/l

Reference substance (positive control):

yes

Remarks:

, potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

Behavioural abnormalities: no
- Immobility of control: no immobility was observed.
- Other adverse effects control: no

Results with reference substance (positive control):

The 48h-EC50 was 0.43 mg/l with a 95% confidence interval between 0.39 and 0.48 mg/l.
The actual responses in this reference test with K2Cr2O7 are within the ranges of the expected responses at the different concentrations, i.e. the 48h-EC50 was between 0.3 and 1.0 mg/l. Hence, the sensitivity of the daphnia was within the range determined with the historical data collected at WIL Research Europe.

Reported statistics and error estimates:

The 24 and 48h-EC50 values could not be determined because the observed effects were below 50%.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 48h-EC50 was >100 mg/l based on analytically confirmed nominal concentrations.