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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
key study
Study period:
18 Oct - 02 Nov 1994
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
no guideline followed
Principles of method if other than guideline:
- Principle of test: Determination of toxic effects of the test item to anaerobic bacteria through reduction of gas production compared to untreated control
- Short description of test conditions:
- test item: Remazol-Brillantgrün 6B
- test duration: 7 days
- temperature 37°C +/- 1°C
- concentrations of the test item: 700 mg/L to 1000 mg/L with ech 3 replicates
- reference item: Dodecyldimethylbenzylammoniumchlorid 50, 100, 200 mg/L
- test vessels: 100 mL bottles filled with 30 mL test medium
- Parameters analysed / observed: injection of phosphoric acid (85%) 30 minutes before measuring the CO2 pressure in the headspace of the bottles..
GLP compliance:

Test material

Constituent 1
Chemical structure
Reference substance name:
Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts
EC Number:
EC Name:
Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts
Cas Number:
Molecular formula:
C32 H16 N8 Ni . C8 H11 N O6 S2 . 2 Na
Nickel, [29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32]-, chlorosulfonyl derivs., reaction products with 2-[(4-aminophenyl)sulfonyl]ethyl hydrogen sulfate monosodium salt, sodium salts
Test material form:

Sampling and analysis

Analytical monitoring:

Test solutions

Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution: 1000 mg test item to 450 mL, pH adjusted from 4,7 to 7,2, stirred for 1,5 hours at 22°C, afterwards pH adjusted from 6,9 to 7,3, stock solution finally filled to 500 mL (concentration: 2000 mg/L). clear solution without undissolved test item
- Controls: yes: test vessel with inoculum, without test item
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): no vehicle
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): -
- Evidence of undissolved material (e.g. precipitate, surface film, etc.): no undissolved material

Test organisms

Test organisms (species):
activated sludge, domestic
Details on inoculum:
- Laboratory culture: no
- Name and location of sewage treatment plant where inoculum was collected: anerobic activated sludge from domestic waste water treatment plant Kriftel
- Preparation of inoculum for exposure: after pretreatment, addition of nutrition solution according to ISO 11734
- Pretreatment: 9 days at 35°C without additing of nutrient solution
- Initial biomass concentration: concentration of activated sludge in test vessels: 3,5 g/L (dry weight)

Study design

Test type:
Water media type:
Limit test:
Total exposure duration:
9 d

Test conditions

Test temperature:
37°C +/- 1°C
Nominal and measured concentrations:
nominal concentrations 700 mg/L and 1000 mg/L
Details on test conditions:
- Test vessel: 100 mL bottles
- Material, size, headspace, fill volume: fill volume: 30 mL
- No. of vessels per concentration (replicates): 3
- Sludge concentration (weight of dry solids per volume): 3.5 g/L (dry weight)
Reference substance (positive control):
Dodecyldimethylbenzyl ammonium chloride

Results and discussion

Effect concentrations
Key result
7 d
Dose descriptor:
other: EC20
Effect conc.:
> 1 000 mg/L
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
inhibition of anaerobic gas production

Any other information on results incl. tables

 concentration [mg/L]  inhibition [%]
 1000  10
 700  6

Applicant's summary and conclusion

Validity criteria fulfilled:
not specified
The 7-d EC20 to anaerobic bacteria was determined >1000 mg/L (nominal) based on anaerobic gas production.