Cuprate(4-), [C-(aminosulfonyl)-C-[[[2-[[4-chloro-6-[(2,5-disulfophenyl)amino]-1,3,5-triazin-2-yl]amino]ethyl]amino]sulfonyl]-29H,31H-phthalocyanine-C,C-disulfonato(6-)-N29,N30,N31,N32]-, sodium salts

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1 September 1993.

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Version / remarks:

OECD Guidelines for Testing of Chemicals (1984) No.209 ''Activated Sludge, Respiration Inhibition Test"

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

EU Method C.11 (Biodegradation: Activated Sludge Respiration Inhibition Test)

Version / remarks:

EEC Commission Directive 87/ 302/ EEC.

Deviations:

not specified

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

No further details specified in the study report.

Analytical monitoring:

no

Details on sampling:

The concentration, homogeneity and stability of the test material preparations were not determined by analysis.

Vehicle:

no

Details on test solutions:

Direct dispersion in water

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

Type: A mixed population of activated sewage sludge micro-organisms.
Source: The aeration stage of the Severn Trent Water Plc sewage treatment plant at Belper, Derbyshire, treating predominantly domestic sewage.
Date of collection: 1 September 1993.
Preparation: The sample was maintained at 21 °C with continuous aeration and used on the day of collection.
The pH was equal to 7.3 and the suspended solids equal to 3.7 g/l prior to use.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

3 h

Post exposure observation period:

No post exposure observation period specified in the study report.

Hardness:

approximately 100 mg/l as CaC03.

Test temperature:

21 °C

pH:

7.3

Dissolved oxygen:

Not applicable - respiration inhibition test

Salinity:

Not applicable - freshwater study

Conductivity:

Not specified

Nominal and measured concentrations:

1000 mg/l nominal - The concentration of the test material preparations was not determined by analysis.

Details on test conditions:

Contact time: 30 minutes and 3 hours.
Vessels: 500 ml conical flasks.
Aeration: Compressed air via narrow bore glass tubes.
Measuring apparatus: 250 ml darkened glass BOD bottle fitted with a Yellow Springs BOD probe, (Model 54) and a magnetic stirrer.
Test concentrations: Test material: 1000 mg/1 in triplicate. Reference material: 3.2, 10 and 32 mg/1.
Temperature: 21 °C
Photoperiod: Normal Laboratory lighting.
Control: Inoculated medium without test or reference material.
Procedure: At time "0" 16 ml of synthetic sewage was made up to 300 ml with water in a 500 ml conical flask. 200 ml inoculum were added and the mixture aerated at approximately 0.5-1 litre per minute. Thereafter, at 15 minute intervals the procedure was repeated with appropriate amounts of the test or reference material being added. As each vessel reached 30 minutes contact time the contents were poured into the measuring vessel and the rate of respiration measured. The procedure was repeated after a 3-hour contact period.

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol.

Key result

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

Details on results:

The rate of respiration for each culture was measured for a 10-minute period (between approximately 8.0 mg O2/l and 1.0 mg O2/l) after 30 minutes and 3 hours contact time.
Percentage inhibition values as calculated from the equation:

% inhibition = [1 – (2RS/{Rc1 + Rc2})] x 100

Where
RS = Oxygen consumption rate for test or reference samples.
Rc1 + Rc2 = Oxygen consumption rates for Controls 1 and 2.

Percentage inhibition is plotted against concentration for the reference material only and the EC50 values derived by inspection for 30 minutes and 3 hours contact time.

JPR Blue 100 EC50 (30 minutes): >1000 mg/l
Variation in respiration rates of Controls 1 and 2: ± 2%

JPR Blue 100 EC50 (3 hours): >1000 mg/L
Variation in respiration rates of Controls 1 and 2: ± 3%

It was considered unnecessary and unrealistic to test at concentrations in excess of 1000 mg/l.

Results with reference substance (positive control):

3,5-dichlorophenol EC50 (3 hours): 10 .5 mg/l

Oxygen Consumption Rates and Percentage Inhibition for 30 Minutes Contact Time for 3,5-dichlorophenol and JPR Blue 100 in an Activated Sludge Respiration Inhibition Test

Concentration (mg/l)	O2consumption rates (mg O2/l/min)	% inhibition
Control            R1                        R2	0.50 0.52	- -
JPR Blue 100            1000    R1            1000    R2            1000    R3	0.52 0.53 0.50	<2> <4> 2
3,5-dichlorophenol                        3.2                        10                        32	0.54 0.29 0.12	<6> 43 76

<increase>

R₁– R₃= Replicates 1 – 3

 

Oxygen Consumption Rates and Percentage Inhibition for 3 Hours Contact Time for 3,5-dichlorophenol and JPR Blue 100 in an Activated Sludge Respiration Inhibition Test

Concentration (mg/l)	O2consumption rates (mg O2/l/min)	% inhibition
Control            R1                        R2	0.44 0.47	- -
JPR Blue 100            1000    R1            1000    R2            1000    R3	0.45 0.48 0.44	1 <5> 3
3,5-dichlorophenol                        3.2                        10                        32	0.39 0.22 0.11	14 52 76

<increase>

R₁– R₃= Replicates 1 - 3

Validity criteria fulfilled:

not specified

Conclusions:

EC50 (3 hours): >1000 mg/l

Executive summary:

TEST TYPE: Activated sludge, respiration inhibition test.

 

TEST METHOD: OECD Guideline No 209 and EEC Commission Directive 87/ 302/ EEC.

 

INOCULUM: Activated sludge micro-organisms.

 

CONTACT TIME: 30 minutes and 3 hours.

 

TEMPERATURE: 21 °C

 

RESULTS

EC50 (30 minutes): >1000 mg/l

EC50 (3 hours): >1000 mg/l

It was considered unnecessary and unrealistic to test at concentrations in excess of 1000 mg/l.

Description of key information

EC50 (3 hours): >1000 mg/l

Key value for chemical safety assessment

EC50 for microorganisms:

1 000 mg/L

Additional information

TEST TYPE: Activated sludge, respiration inhibition test.

TEST METHOD: OECD Guideline No 209 and EEC Commission Directive 87/ 302/ EEC.

INOCULUM: Activated sludge micro-organisms.

CONTACT TIME: 30 minutes and 3 hours.

TEMPERATURE: 21 °C

 

RESULTS

EC50 (30 minutes): >1000 mg/l

EC50 (3 hours): >1000 mg/l

It was considered unnecessary and unrealistic to test at concentrations in excess of 1000 mg/l.