2,3-epoxypropyltrimethylammonium chloride

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003-01-06 to 2003-01-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to the appropriate OECD test guideline, and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method C.3 (Algal Inhibition test)

Version / remarks:

Cited as Directive 92/69/EEC, (1992)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Alga, Growth Inhibition Test)

Version / remarks:

(1984)

Deviations:

yes

Remarks:

lux range is higher than guideline, no preliminary test was conducted, initial cell concentration was 2 x 10^4 cells/ml (rather than the guideline recommended 2 x 10^4 cells/ml).

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2,3-epoxypropyltrimethylammonium chloride

EC Number:

221-221-0

EC Name:

2,3-epoxypropyltrimethylammonium chloride

Cas Number:

3033-77-0

Molecular formula:

C6H14NO.Cl

IUPAC Name:

N,N,N-trimethyl(oxiran-2-yl)methanaminium chloride

Details on test material:

- Name of test material (as cited in study report): 2,3-Epoxypropyltrimethylammoniumchloride; QUAB 151; EPTAC

- Physical state: clear liquid

- Stability under test conditions: The stability of the test substance was checked before the main test by measuring the concentrations of the substance, without the algae, after 0 and 72 hours.

- Storage condition of test material: 5 degC

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: A series of concentrations tested was made up for the stability control in the same way as the concentrations for biological part, but without test organisms. Concentrations deviated only slightly (less than 20 %) from each other. Therefore, the main test was performed with nominal concentrations.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION

- Method: A stock solution of 2.56 g/L (ca. 2 g of test iten without water/L) was equilibrated in deionised water.

Test organisms

Test organisms (species):

Desmodesmus subspicatus (previous name: Scenedesmus subspicatus)

Details on test organisms:

TEST ORGANISM

- Strain: 86.81 SAG

- Source (laboratory, culture collection): Albrecht von Hailer- Institut fur Pflanzenwissenschaften of the University of Gottingen, Abteilung Experimentelle Phykologie, Nikolausberg 18, Gottingen, Germany, subsequently in-house breeding.

- Age of inoculum (at test initiation): 3 days


ACCLIMATION

- Culturing media and conditions (same as test or not): same as test

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

72 h

Test conditions

Test temperature:

range:  22.9 - 23.4 °C

pH:

At the beginning of the test: 7.7 - 7.8. At the end of the test: 8.2 - 9.7. It was stated in the test report that due to the strong growth and the relative low concentration of buffer reagent, in some of the test concentrations and in the controls an increase of pH was observed. Since the growth was not influenced by this effect, it was not regarded as important for the quality of the test.

Nominal and measured concentrations:

Nominal concentrations: 0, 70, 120, 200, 340, 580, 1000 mg/L.
It was stated in the test report that the concentrations were based on the test item without water.

Details on test conditions:

TEST SYSTEM

- Test vessel:

- Material, size, headspace, fill volume: Erlenmeyer flasks

- Aeration: yes

- Initial cells density: 2 x 10^4 cells/ml

- Control end cells density: 142 x 10^4 cells/ml

- No. of vessels per concentration (replicates): 3

- No. of vessels per control (replicates): 3


GROWTH MEDIUM

- Standard medium used: yes



TEST MEDIUM / WATER PARAMETERS

- Source/preparation of dilution water: deionised water

- Culture medium different from test medium: no


OTHER TEST CONDITIONS

- Sterile test conditions: yes

- Light intensity and quality: 6000-10000 lux, white


EFFECT PARAMETERS MEASURED:

- Determination of cell concentrations: cell numbers were determined photometrically. Therefore, the absorption was measured at 685 nm and the measurements were converted by means of a calibration curve into the parameter "cell numbers/ml". Cell numbers were evaluated every 24 h


TEST CONCENTRATIONS

- Spacing factor for test concentrations: ~1.7

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

580 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: cell growth

Remarks on result:

other: test. mat. based on test item excluding water

Effect concentrations 2

Duration:

72 h

Dose descriptor:

other: ErC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Biomass and growth rate

Remarks on result:

other: test. mat. based on test item excluding water

Effect concentrations 3

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

814 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth rate

Remarks on result:

other: test. mat. based on test item excluding water

Effect concentrations 4

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

625 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks on result:

other: test. mat. based on test item excluding water

Details on results:

- Exponential growth in the control (for algal test): yes

Reported statistics and error estimates:

EC10 and NOEC values were determined graphically, the EC50 was set as a default to > highest concentration, as 100% mortality was not achieved.

Any other information on results incl. tables

Table 1. Summary of findings.

 

Nominal concentration (mg/L)	Average number of cells (1 x 10^4 cells/ml)				Biomass inhibition (%)	Growth rate inhibition (%)
	0h	24h	48h	72h
Control	2	11	30	142	0	0
70	2	11	27	141	3.3	0.1
120	2	10	22	123	17.3	3.4
200	2	11	29	136	3.7	1.0
340	2	10	25	120	15.9	3.9
580	2	11	29	134	4.7	1.3
1000	2	8	21	74	43	15.3

 

Initial cell concentration is reported as 7.38 x 10^4 cell/ml in the control and ~8 x 10^4 in the other treatment levels.

 

 

Table 2. Measured concentrations of test item in solution at 0 and 72 h without test organism.

 

Nominal concentration (mg/L)	0 h sample (mg/L)	72 h sample (mg/L)
70	75	69
120	122	117
200	211	200
340	351	339
580	575	584
1000	1018	1004
Stock solution (2000)	1918	-

 

The measured concentrations after 72 h did not deviate by more than 20% from the freshly prepared test solutions. The measured recovery rates (nominal values compared with the analytical values after 0 h) amounted to >80%.

The concentrations refer to the test item with the exclusion of water.

 

Applicant's summary and conclusion

Validity criteria fulfilled:

no

Remarks:

pH shifted by more than 1 unit. Thought to have not adversely affected the results of the test.

Conclusions:

A 72 h ErC50 value of >1000 mg (test item without water)/L and a NOEC value of 580 mg/L have been determined for the effects of the test substance on growth rate of the algae D. subspicatus.