1-Octadecanaminium,N,N-Dimethyl-N-Octadecyl-,(Sp-4-2)-[29h,31h-Phthalocyanine2-Sulfonato(3-)-N 29,N 30,N 31,N 32]Cuprate(1-)

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study; well-documented study report

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2001

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Details on test material:

- Analytical purity: 81%
- Lot/batch No.: F70305T
- Expiration date of the lot/batch: 2006

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

- Method: The test material was weighed into cups constructed of aluminum foil and the cups an the test substance were added to the respective test vessels which contained reverse osmosis water and synthetic sewage. The vessels were placed in an ultrasonic bath (Decon Fs400b) and their contents subjected to ultrasonification for approximately ten minutes to disperse the test substance in the test medium before adding the inoculum

Test organisms

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

- Method of cultivation: Collected from one of the return lines of Burley Menston sewage treatment works. The sample was aerated by means of a compressed air supply delivered through a diverse block
- Preparation of inoculum for exposure: The inoculum was used in the range-finder two days after the sludge sample was collected. The inoculum was maintained by aeration at all times and was fed overnight with 400 mL synthetic sewage on two occasions during the interval between collection and use.
- Initial biomass concentration: 4 g/L (initials suspended solids concentration was 4.56g/L therefore it was diluted with de-chlorinated tap water to provide 8L inoculum with a nominal solids content of 4 g/L)

Study design

Test type:

other: Direct addition

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test conditions

Test temperature:

19.4 to 21.6 C

pH:

Bulk inoculum pH was 6.09

Nominal and measured concentrations:

1.0, 10, 100, and 1000 mg/L (nominal)

Details on test conditions:

TEST SYSTEM
- Material, size, headspace, fill volume: 500mL total (300 mL of synthetic sewage sludge (16mL) diluted with reverse-osmosis water and 200 mL of activated sludge)
- Aeration: yes
- No. of vessels per concentration (replicates): duplicate
- No. of vessels per control (replicates): unreplicated
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reverse-osmosis water
OTHER TEST CONDITIONS
- Adjustment of pH: used 1M NaOH
TEST CONCENTRATIONS
- Test concentrations: 1.0, 10, 100, and 1000 mg/L
- Results used to determine the conditions for the definitive study: Absence of significant toxic effect up to 1000 mg/L in the range finding test, therefore further testing was considered unnecessary.

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

> 1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: The test substance did not inhibit the respiration of activated sludge at concentrations up to and including 1000 mg/L. The EC50 of the test substance could not be determined, but it exceeded 1000 mg/L.

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: 3,5 dichlorophenol EC50 was 9 mg/L

Any other information on results incl. tables

Treatment	Log10conc	Respiration rate mg O2/Lh	Mean	Standard Deviation σn-1	Percentage Inhibition
Control	na	77.25^1	78.83	2.0	Na
	na	78.25^1
	na	81.04^2
	na	90.11^2,3
1.0 mg/L Test material	0.0000	84.38	83.90	0.7	0
		83.41
10 mg/L Test material	1.0000	85.27	84.94	0.5	0
		84.61
100 mg/L Test material	2.0000	88.47	85.06	4.8	0
		81.64
1000 mg/L Test Material	3.0000	89.27	84.54	6.7	0
		79.80
5mg/L 3,5-DCP	0.6990	53.09	na	na	33
15mg/L 3,5-DCP	1.1761	28.12	na	na	64
45mg/L 3,5-DCP	1.6532	6.28	na	na	92

¹start-of-series control

²end of series control

³outside 15% acceptance limit. Omitted from mean

na: not applicable

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test substance did not inhibit the respiration of activated sludge at concentrations up to and including 1000 mg/L. The EC50 of the test substance could not be determined, but exceeded 1000 mg/L.

Executive summary:

The toxicity to microorganisms of the test material was tested using activated sludge of a predominately domestic sewage according to OECD guideline 209. In the range-finder phase study, doses of 1.0, 10, 100, and 1000 mg/L (nominal) test material was added to the inoculum. The test substance did not inhibit the respiration of activated sludge at any concentrations tested, and further testing was considered unnecessary. Both the OECD guideline validity criteria that related to the reference inhibitory response and conformity of the control respiration rates were met. The results of this study are considered valid. The EC50 of the test substance could not be determined from the results of this study, but it exceeded 1000 mg/L. Based on these results the test article is considered to be non-toxic to microorganisms at the concentrations tested.