3-(D-gluconoylamino)propyl(2-hydroxyethyl)dimethylammonium chloride

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Experimental start date 02 Mar 2017 - experimental completion date 03 Mar 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test (Carbon and Ammonium Oxidation))

Version / remarks:

adopted 22 July 2010

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-(D-gluconoylamino)propyl(2-hydroxyethyl)dimethylammonium chloride

EC Number:

257-440-3

EC Name:

3-(D-gluconoylamino)propyl(2-hydroxyethyl)dimethylammonium chloride

Cas Number:

51812-80-7

Molecular formula:

C13H29N2O7.Cl

IUPAC Name:

1-Propanaminium, 3-(D-gluconoylamino)-N-(2-hydroxyethyl)-N,N-dimethyl-, chloride

Test material form:

liquid: viscous

Details on test material:

CR Test item 207516/A
Identification Quaternium-22
Appearance Brown paste (determined by Charles River Den Bosch)
Batch 0001954895
Purity/Composition > 90%
Test item storage In refrigerator (2-8°C)
Stable under storage conditions until 16 June 2017 (retest date)

Specific details on test material used for the study:

Test item 207516/A
Identification Quaternium-22
Appearance Brown paste (determined by Charles River Den Bosch)
Batch 0001954895
Purity/Composition > 90%
Test item storage In refrigerator (2-8°C)
Stable under storage conditions until 16 June 2017 (retest date)

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

A stock solution of 10 g/L was prepared by adding 4.96565 g test item to 500 mL of Milli-RO water (tap water purified by reverse osmosis; Millipore Corp., Bedford, Mass., USA). Magnetic stirring for 34 minutes was applied to accelerate dissolution and to ensure homogeneity. Volumes of the clear brown/yellow stock solution (pH 5.7) corresponding to the test concentration were then added to the test media. Optimal contact between the test item and test organisms was ensured by applying continuous aeration and stirring.

Test organisms

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

Source: Municipal sewage treatment plant: 'Waterschap Aa en Maas', 's- Hertogenbosch, The Netherlands.
Preparation of the sludge: The sludge was coarsely sieved (1 mm) and allowed to settle. The supernatant was removed and ISO-medium was added. A small amount of the sludge was weighed and dried overnight at ca. 105°C to determine the amount of suspended solids (3.0 g/L of sludge, as used for the test). The pH was 7.7 on the day of testing. The batch of sludge was used one day after collection; therefore 50 mL of synthetic medium (=sewage feed) was added per litre of activated sludge at the end of the collection day. The sludge was kept aerated at test temperature until use
Medium: Adjusted ISO-medium, formulated using RO-water (tap water purified by reverse osmosis; GEON Waterbehandeling, Berkel-Enschot, The Netherlands) with the following composition:
CaCl2.2H2O 211.5 mg/L
MgSO4.7H2O 88.8 mg/L
NaHCO3 46.7 mg/L
KCl 4.2 mg/L
Rationale Recognized by international guidelines as the recommended test system.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

3 h

Test conditions

Test temperature:

ranged between 18 and 21°C

pH:

between 7.2 and 8.3

Dissolved oxygen:

above 60-70% saturation (60% of air saturation is > 5 mg/L at 20°C)

Nominal and measured concentrations:

Combined limit/range-finding: 10, 100 and 1000 mg/L

Details on test conditions:

Vessels All glass open bottles/vessels.
Milli-RO / Milli-RO water Tap water purified by reverse osmosis (Millipore Corp., Bedford, Mass., USA).
Synthetic medium 16 g peptone
(=sewage feed) 11 g meat extract
3 g urea
0.7 g NaCl
0.4 g CaCl2.2H2O
0.2 g MgSO4.7H2O
2.8 g K2HPO4
Dissolved in Milli-RO water, made up to 1 litre and filtered. The pH was within 7.5 ± 0.5.
Air supply Clean, oil-free air.
Performance of the test The synthetic medium (= sewage feed, 16 mL) and an appropriate amount of the test item stock were mixed and made up to 250 mL with Milli-RO water. The pH was determined. Thereafter, 250 mL activated sludge was added. This was the start of the test. After the 3-hour contact time the oxygen consumption was recorded for a period of approximately 10 minutes. During measurement, the sample was not aerated but continuously stirred on a magnetic stirrer.
pH: The pH was determined in the remaining part of the reaction mixture. This procedure was repeated for all test/reference item concentrations and controls.
Temperature: the medium temperature was recorded continuously in two temperature control vessels. The temperature control vessels were identically prepared compared to the control vessels. A temperature control vessel with a REES sensor was placed in each fume cupboard of the climate room.
Oxygen recording: Determination of oxygen was performed with multiple oxygen sensors connected to a BlueBox (GO-Systemelektronik GmbH, Germany), a multichannel measuring and controlling system.

Reference substance (positive control):

yes

Remarks:

3,5- dichlorophenol: 1.0, 3.2, 10 and 32 mg/L.

Results and discussion

Effect concentrations

Effect concentrations 1

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

Effect concentrations 2

Key result

Duration:

3 h

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the conditions of this present test Quaternium-22 was not toxic to waste water bacteria (activated sludge) at 1000 mg/L.
The EC50 was above 1000 mg/L.

Executive summary:

The influence of Quaternium-22 on the respiration rate of activated sludge was investigated after a contact time of 3 hours.The study procedures described in this report were based onthe OECD guideline No. 209, 2010.

The batch of Quaternium-22 tested was a brown paste (UVCB) with a purity of > 90% (water content <10%).No correction was made for the purity/composition of the test item.

A stock solution of 10 g/L was prepared. Magnetic stirring for 34 minutes was applied to accelerate dissolution and to ensure homogeneity.Volumes of the clear brown/yellow stock solution corresponding to the test concentration were then added to the test media. Optimal contact between the test item and test organisms was ensured by applying continuous aeration and stirring. Thereafter, oxygen consumption was recorded for approximately 10 minutes.

In a combined limit/range-finding test concentrations of 10, 100 and 1000 mg/L were tested. The highest concentration was tested in triplicate; lower concentrations consisted of one replicate. Furthermore, at 1000 mg/L an abiotic control (1 replicate) was tested.Responses were compared to the blank.

No statistically significant inhibition of the respiration rate of the sludge was recordedat a concentration of 1000 mg Quaternium-22 per litre.Therefore, the EC₅₀was above the highest concentration tested (1000 mg/L).