Reaction mass of 29H,31H-phthalocyaninato(2-)-N29,N30,N31,N32 copper and hydrogen [29H,31H-phthalocyaninesulphonato(3-)-N29,N30,N31,N32]cuprate(1-), compound with dodecylamine (1:1)

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:

04 February 2016 - 04 February 2016

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 (Carbon and Ammonium Oxidation))

Deviations:

no

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Storage condition: Controlled room temperature (15-25°C, below 70 RH%)

Analytical monitoring:

not specified

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: Because of the low solubility, the test item was added by directly weighing the test item into the bottles.

- Controls: Abiotic Control, untreated control, and reference control

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

- Name and location of sewage treatment plant where inoculum was collected: Veszprém, Hungary

- Preparation of inoculum for exposure: The activated sludge used for this study was washed and centrifuged and the supernatant liquid phase was decanted. The solid material was re-suspended in chlorine-free tap water and again centrifuged. An aliquot of the final sludge suspension was weighed, dried and the ratio of wet sludge to dry weight determined.
Based on this ratio, calculated amounts of wet sludge were suspended in chlorine-free tap water to yield a concentration equivalent to 3 g per litre (on dry weight basis). The pH of the activated sludge inoculum was determined to be pH 7.43. The activated sludge was used directly after conditioning.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Remarks on exposure duration:

n/a

Post exposure observation period:

n/a

Hardness:

not specified

Test temperature:

20.8 - 21.4°C

pH:

7.31 - 8.29

Dissolved oxygen:

A well-mixed sample of each treatment was poured into a BOD flask after about 3 hours incubation time, and was not further aerated. The oxygen concentration was measured for about fifteen minutes, with an O2 electrode and was recorded in every half minute. The oxygen consumption (in mg O2 L-1 minute-1) was determined from the most linear part of the respiration curve.

Salinity:

not specified

Conductivity:

not specified

Nominal and measured concentrations:

Test groups (nominal concentration): 10, 100, and 1000 mg/L
Abiotic Control: 1000 mg/L nominal concentration
Reference item: 1.0, 3.2 and 10.0 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: glass flask
- Material, size, headspace, fill volume: glass beakers for 500 mL volume and BOD bottles with 300 mL volume
- Aeration: With compressed air (about 2 L/min)
- No. of vessels per control (replicates): 5
- No. of vessels per vehicle control (replicates): 5
- No. of vessels per abiotic control (replicates): 5
- Sludge concentration (weight of dry solids per volume): 3 g per litre
- Nutrients provided for bacteria: daily with 50 mL synthetic sewage

Reference substance (positive control):

yes

Remarks:

3,5-Dichlorophenol (3,5-DCP)

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:

In comparison to the inoculum controls the inhibition of the respiration rate in the case of the activated sludge was between 7.10 % and 40.13 % in the examined range of 10 – 1000 mg/L of B331 test item.
The average specific respiration rate (RS) of the untreated (Blank) controls (B1-B5) was 35.05 mg O2/L/h/g solid.
The variation coefficient of oxygen uptake rate in above untreated replicates was 6.41 %.

Results with reference substance (positive control):

- Results with reference substance valid? yes
- Relevant effect levels: 3-hour EC50 of 3,5-Dichlorophenol was calculated to be 12.51 mg/L

Reported statistics and error estimates:

not specified

Validity criteria fulfilled:

not specified

Conclusions:

Under the conditions of this Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation) the observed endpoint for the effect of B331 was the following:
The 3-hour EC50: > 1000 mg/L
Based on the results of this study, the test item B331 had no inhibition effect on the total respiration rate of the microorganisms of the activated sludge, therefore further testing for nitrification inhibition is not necessary.

Description of key information

EC50 (3 hours) > 1000 mg/L

Additional information

An Activated Sludge Respiration Inhibition Test (ASRIT) was conducted (CiToxLAB Hungary, 2016) to assess the short-term effects of B331 on Sewage Treatment Plant microorganisms. The study was conducted according to OECD test guideline 209 and EC method C11, and in compliance with GLP. The inhibitory effect of the test material was assessed at concentrations 10, 100 and 1000 mg/L over 3 hours.

Less than 50% inhibition of respiration was observed at the highest test concentration (3 hour EC50 > 1000 mg/L); on this basis the test material was considered to be non-inhibitory to STP microorganisms.