Benzyldimethyl[2-[(1-oxoallyl)oxy]ethyl]ammonium chloride

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:

toxicity to aquatic algae and cyanobacteria

Adequacy of study:

other information

Study period:

04/10/2010-08/11/2010

Rationale for reliability incl. deficiencies:

other: Well documented report of a guideline study conducted to GLP.

Data source

Reference

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test material

Test material information

Constituent 1

Reference substance name:

Automatically generated during migration to IUCLID 6, no data available

IUPAC Name:

Automatically generated during migration to IUCLID 6, no data available

Details on test material:

- Name of test material (as cited in study report): ADAMQUAT BZ
- Trade name: Flocryl ADAM / BZ CL 80
- Physical state: light yellow liquid.
- Analytical purity: 80%
- Impurities (identity and concentrations): Water = 20%, EMHQ (Stabiliser) < 500 ppm, acrylic acid < 100 ppm, benzyl chloride < 100 ppm
- Composition of test material, percentage of components: 100%
- Purity test date:09/11/10
- Lot/batch No.: AD/20021980
- Expiration date of the lot/batch: 24/11/10
- Stability under test conditions: at room temperature and away from the light
- Storage condition of test material: stable

Sampling and analysis

Details on sampling:

In order to verify that initial concentrations were equivalent to nominal concentrations (soluble, stable substances) chemical analysis was performed at the beginning of the definitive test and prior to addition of the organisms by sampling test solutions. Final concentrations were measured at the end of the test period (as described in OECD 201 paragraph 37).

Test solutions

Details on test solutions:

For the range finding test, a stock solution at 100 mglL was prepared 30 min. before the beginning of the test by mixing 100 mg of the test item in 1 litre of dilution water. It was used to prepare the convenient range of concentrations: 100, 50, 10, 5, 1 and 0.1 mg/L.
For the definitive test a stock solution at 50 mglL was prepared the same way 45 min. before the beginning of the test and used to prepare the convenient range of concentrations: 50, 26, 13.5, 7, 3.7, 1.9 and 1 mg/L.

Test organisms

Details on test organisms:

TEST ORGANISM
- Common name: Pseudokirchneriella subcapitata (previously known as Raphidocelis subcapitata and Selenastrum capricornutum)
- Strain: CCAP 278/4 stock
- Source: Culture Centre of Algae and Protozoa (Ambleside, UK)
- Age of inoculum (at test initiation): i week
- Method of cultivation: The conditions used for culturing algae are described in Annex 2 of the OECD 201 guideline. Two flasks, each containing approximately 100 mL of axenic stock culture of algae were incubated at 23±1°C under lighting (photoperiod: 16 hours of illumination, 8 hours of darkness), shaken slowly and continuously. These stock cultures were renewed every week, using two new cultures. The quality of the stock culture was verified for the absence of micro-organisms and deformed cells under microscopic observation before use.
Three days before the beginning of the study two pre-cultures were prepared by inoculating each stock suspension of algae (5 mL) into sterile dilution water (500 mL). The pre-cultures were incubated under the same conditions as those used for the stock cultures. Only one of the two pre-cultures was used to inoculate the test flasks; the second one was to be used only if the first one was damaged.
ACCLIMATION
- Acclimation period: none required
- Culturing media and conditions (same as test or not): same
- Any deformed or abnormal cells observed: no

Study design

Total exposure duration:

72 h

Post exposure observation period:

None required

Test conditions

Hardness:

50 mg/l (NaHCO3)

Test temperature:

23 ± 1°C

pH:

At T0: 7.34 - 8.34

Dissolved oxygen:

At T0: 8.4-8.6 mg/l

Salinity:

Not specified

Nominal and measured concentrations:

Nominal: 0, 1, 1.9, 3.7, 7, 13.5, 26 and 50 in the definitive test (0, 0.1, 1, 5, 10, 50 and 100 mg/l in the preliminary test)
Measured: 0, 1, 2.2, 3.9, 5.8, 14.7, 28 and 53 mg/l in the definative test

Details on test conditions:

TEST SYSTEM
- Test vessel: 120 ml glass flask
- Type (delete if not applicable): closed
- Material, size, headspace, fill volume: 50 ml
- Aeration: none
- Initial cells density: 1.28E+06 cells/ml
- Control end cells density: 1.17E+06
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6
GROWTH MEDIUM
- Standard medium used: yes
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ultra pure (resistivity = 18 MΩ)
- Culture medium different from test medium: no
- Intervals of water quality measurement: beginning and end of test
OTHER TEST CONDITIONS
- Sterile test conditions: yes
- Adjustment of pH: no
- Photoperiod: 16 hours light/8 hours dark
- Light intensity and quality: constant illumination by 8 fluorescent tubes between 6,000 and 10,000 lux (Mazdafluor, white industry 33)
EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Determination of cell concentrations: cell counter (Beckman Coulter Z2)
TEST CONCENTRATIONS
- Spacing factor for test concentrations: 1.92 (based on the results of preliminary test the 7 concentrations tested were: 50, 26, 13.5, 7, 3.7, 1.9 and 1 mg/L)
- Range finding study
- Test concentrations: 100, 50, 10, 5, 1 and 0.1 mg/L
- Results used to determine the conditions for the definitive study: 50, 10, 5, 1 and 0.1 mg/L

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

72 h

Effect conc.:

5.05 mg/L

Effect concentrations 2

Duration:

72 h

Effect conc.:

14 mg/L

Remarks on result:

other: 8.1-24

Effect concentrations 3

Duration:

72 h

Effect conc.:

21 mg/L

Remarks on result:

other: 15-34

Details on results:

- Exponential growth in the control (for algal test): yes
- Observation of abnormalities (for algal test): no
- Unusual cell shape: no
- Colour differences: no
- Flocculation: no
- Adherence to test vessels: no
- Aggregation of algal cells: no
- Any stimulation of growth found in any treatment: no
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: no
- Effect concentrations exceeding solubility of substance in test medium: no

Results with reference substance (positive control):

Not applicable

Reported statistics and error estimates:

The biomass in the control cultures increased exponentially by a factor of 161 (corresponding to a specific growth rate of 1.694 day-1) within the 72-hour test period, which is higher than the minimal value (R = 16) mentioned OECD guideline 201.

Any other information on results incl. tables

Nominal and measured concentrations of the test itemat the beginning and at the end ofthe exposure period

 

Concentration ofADAMQUAT BZ
Nominal	Measured in non inoculated solutions			Extrapolated*
	Initial	Final **	Final/Initial
(mg/l)	(mg/l)	(mg/l)	%	(mg/l)
50	53	34	64.2	42.45
26	28	18 (18)	64.3	22.45
13.5	14.7	9.6 (1.4)	65.3	11.88
7	5.8	4.4 (< DL)	75.9	5.05
3.7	3.9	2.5 (< DL)	64.1	3.12
1.9	2.2	1.2	54.5	1.62
1	1.0	< DL	-	0.32
0	< DL	< DL	-	-

 

< DL : concentration lower than the Detection Limit of the analytical method (0.2 mg/L).

< QL : concentration lower than the Quantification Limit of the analytical method (0.7 mg/L).

* Extrapolated concentration with geometric average value of initial and final concentrations. When final concentration is lower than DL or QL, half the values of DL or QL are used for the calculation.

** Within parenthesis: concentrations of ADAMQUAT BZ measured in inoculated series (with algae).

 

Applicant's summary and conclusion

Conclusions:

The test substance, ADAMQUAT BZ, was harmful tothe freshwater algae Pseudokirchneriella subcapitata after 72 hours in a static test. The EC50 (biomass) was determined to be 14 mg/l and the NOEC (biomass and growth rate) was 5.05 mg/l.