Reaction mass of 3-[(2-aminoethyl)ammonio]propane-1-sulfonate and 3,3'-(ethane-1,2-diyldiammonio)dipropane-1-sulfonate and ethane-1,2-diamine

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

June 2008

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-[(2-aminoethyl)amino]propane-1-sulfonic acid

Cas Number:

14235-54-2

Molecular formula:

C5H14N2O3S

IUPAC Name:

3-[(2-aminoethyl)amino]propane-1-sulfonic acid

Details on test material:

- Physical state: liquid
- Composition of test material, percentage of components:
48.5 % (w/w) 3-[(2-aminoethyl)ammonio]propane-1-sulfonate;
51.5 % water
- Lot/batch No.: 06060423/101
- Stability under test conditions: stable
- Storage condition of test material: room temperature in the dark

Sampling and analysis

Analytical monitoring:

not required

Test organisms

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

For the preparation of the microbial inoculum activated sludge was used. The sludge was collected from one of the return sludge channels of the sewage treatment plant in Baden near Vienna (Zubringerstraße, A-2500 Baden). Baden is a spa with only a small amount of industry and the waste water of the sewage treatment plant is predominantly domestic.

A sample of activated sludge was collected one day before the test. The container for transportation was only maximally 2/3 filled in order to maintain the contact with air.

At arrival in the laboratory (1 hour after sample collection), the supernatant over the sludge was decanted, the sludge was diluted with tap water and aerated by means of compressed air. 50 mL of synthetic sewage concentrate were added per litre each day. On the day of the test the sludge was allowed to sediment and the supernatant was decanted.
The concentration of suspended solids was determined by filtering 3 mL of the sample through a pre-dried and pre-weighed glass microfibre filter. After drying at 105 °C and re-weighing the amount of solids was calculated.

On the basis of this calculation the concentration of suspended solids was adjusted to 4 g/L with tap water. This suspension was used as inoculum for the samples. The inoculum was aerated until use.

Initial biomass concentration: 4 g/L

Study design

Test type:

static

Water media type:

brackish water

Total exposure duration:

4 h

Test conditions

Test temperature:

20 °C

pH:

conc. 1 (7.7 mg/L) 8.27 at the end
conc. 2 (19.2 mg/L) 8.27 at the end
conc. 3 (48.1 mg/L) 8.24 at the end
conc. 4 (120.2 mg/L) 8.27 at the end
conc. 5 (300.5 mg/L) 8.26 at the end

Dissolved oxygen:

conc. 1 (7.7 mg/L) 6.44 mg/L O2 at the start, 2.50 mg/L O2 at the end
conc. 2 (19.2 mg/L) 6.19 mg/L O2 at the start, 2.12 mg/L O2 at the end
conc. 3 (48.1 mg/L) 5.98 mg/L O2 at the start, 1.92 mg/L O2 at the end
conc. 4 (120.2 mg/L) 5.92 mg/L O2 at the start, 2.34 mg/L O2 at the end
conc. 5 (300.5 mg/L) 6.62 mg/L O2 at the start, 2.92 mg/L O2 at the end

Nominal and measured concentrations:

Nominal 7.7, 19.2, 48, 120 and 300 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: open 1 L glass beakers, aeration with 0.7 L air / minute with a pasteur pipette.
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2
- No. of vessels per vehicle control (replicates): 3
- Biomass loading rate: 1.6 g dry material / L.


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: tap water.


OTHER TEST CONDITIONS
- Adjustment of pH: no


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Oxygen concentration.


TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2.5
- Range finding study: no

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Results and discussion

Effect concentrations

Duration:

4 h

Dose descriptor:

EC50

Effect conc.:

> 300 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Remarks:

(water free)

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: EC50 = 14.3 mg/L

Any other information on results incl. tables

Components and pH of the samples

sample	stock solution, concentr. (mg/L)	stock solution, amount (mL)	tap water (mL)	synthetic sewage solution (mL)	adjusted microbial inoculum (mL)	final concentr. (mg/L)	pH at the end of the incubation
neg. control 1	0	0	100	200	200	0.0	8.26
test substance 1	1503	2.56	ad 100	200	200	7.7	8.27
test substance 2	1503	6.4	ad 100	200	200	19.2	8.27
test substance 3	1503	16	ad 100	200	200	48.1	8.24
test substance 4	1503	40	ad 100	200	200	120.2	8.27
test substance 5	1503	100	0	200	200	300.5	8.26
neg. control 2	0	0	100	200	200	0.0	8.23
DCP 1	523	5.0	ad 100	200	200	5.2	8.22
DCP 2	523	12.2	ad 100	200	200	12.8	8.29
DCP 3	523	30.0	ad 100	200	200	31.4	8.29

Respiration rate and inhibition of the samples

sample	final concentr. (mg/L)	linear range (minutes)	mg O2/ L at begin	mg O2/ L at end	respiration rate (mg O2. L^-1. h^-1)	percent respiration inhibition
neg. control 1	0.0	8.44	5.65	1.72	27.94	5.99
test substance 1	7.7	8.52	6.44	2.50	27.75	6.64
test substance 2	19.2	8.48	6.19	2.12	28.80	3.10
test substance 3	48.1	7.96	5.98	1.92	30.60	-2.97
test substance 4	120.2	7.36	5.92	2.34	29.18	1.80
test substance 5	300.5	8.00	6.62	2.92	27.75	6.63
neg. control 2	0.0	8.00	5.99	1.79	31.50	-5.99
DCP 1	5.2	8.12	5.86	2.33	26.08	12.23
DCP 2	12.8	9.12	7.53	5.37	14.21	52.18
DCP 3	31.4	9.12	7.82	6.66	7.63	74.32

Applicant's summary and conclusion

Validity criteria fulfilled:

yes