Registration Dossier

Registration Dossier

Diss Factsheets

Ecotoxicological information

Toxicity to microorganisms

Currently viewing:

Administrative data

Link to relevant study record(s)

Referenceopen allclose all

Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
February 22nd, 2010 to February 24th, 2010
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment it is considered appropriate to read across from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the structurally analogous quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

Furthermore, in certain organic solvents it has been reported that the exchange constants between nitrite and chloride in quaternary ammonium salts (QAS) are approximately equal. [Zhurnal Analiticheskoi Khimii, 2010, Vol. 65, No. 6, pp. 579–584. (E.M. Rakhman’ko, M.S. Markovskaya, L.S. Stanishevskii, Yu.S. Zubenko, A.R. Tsyganov)]

In this study Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1) was subjected to a toxicity to microorganisms (activated sludge, respiration inhibition test) study according to OECD Guideline 209.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test
Deviations:
no
GLP compliance:
yes
Specific details on test material used for the study:
- Name of test material (as cited in study report): Quaternary ammonium compounds, di-C12-18 alkyldimethyl chlorides
- Physical state: Light yellowish paste
- Analytical purity: 99.6 %
- Lot/batch No.: 10-SK005
- Expiration date of the lot/batch: Jan 2012
- Stability under test conditions: Stable under test conditions
- Storage condition of test material: At ambient temperature in the dark
Analytical monitoring:
not required
Details on sampling:
No data
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution was prepared by stirring 0.46 g quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides in 460 mL of deionized water. The stock solution was a stable homogenous suspension (very light turbidity).
- pH stock solution = 5.0
Test organisms (species):
activated sludge of a predominantly domestic sewage
Details on inoculum:
- Laboratory culture: Unadapted secondary activated sludge was obtained (22-02-2010) from the WWTP Nieuwgraaf in Duiven. The WWTP Duiven is an activated sludge plant treating predominantly domestic wastewater.
- Method of cultivation: Prior to use the activated sludge was homogenized with a syringe. The dry weight of the
homogenized activated sludge was determined and subsequently concentrated by settlement to the required dry weight concentration.
- Preparation of inoculum for exposure: During the test period the activated sludge was kept in a thermostated 2 L cylindrical glass vessel to maintain the temperature at 20 ± 2 °C. A magnetic stirrer bar at the bottom and a glass frit were used to agitate and aerate the activated sludge. The dry weight of the activated sludge in the incubation vessels was 1.7 g/L.
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
3 h
Post exposure observation period:
No data
Hardness:
No data
Test temperature:
20.2- 21.0 °C
pH:
6.9- 7.2
Dissolved oxygen:
No data
Salinity:
No data
Nominal and measured concentrations:
Nominal: 0, 10, 20, 40, 80, 160 and 320 mg/L
Details on test conditions:
- The dry weight concentration of the activated sludge in the test was 1.7 g/L
- The inhibitory effect of the test substance/reference compound is expressed as a percentage of the mean of the two control respiration rates.

TEST SYSTEM
- Test vessel: 250- 300 mL Erlenmayers

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap-water
- The homogenized activated sludge was incubated in a shaking water bath (100 rpm, 20°C) for 3 hours with various concentrations of the test compound and synthetic sewage.
- Synthetic sewage was added to the activated sludge (1.6 mL on a total volume of 50 mL).
Synthetic sewage contained per liter: 16 g peptone
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
Test conditions
The pH of the reaction mixtures after the incubation period ranged from 6.9 to 7.2. Temperature varied from 20
Reference substance (positive control):
yes
Remarks:
3,5-dichlorophenol
Key result
Duration:
3 h
Dose descriptor:
EC50
Effect conc.:
68 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
inhibition of total respiration
Remarks on result:
other: with 95% confidence limits of 15 and 858 mg/L.
Details on results:
The EC50 of test material for activated sludge after 3 h contact time is 68 mg/L with 95% confidence limits of 15 and 858 mg/L. The EC10, EC20 and EC80 are 12, 22 and 215 mg/L, respectively
Results with reference substance (positive control):
The EC50 of the reference compound was 8 mg/L which is within the prescribed range of 5 to 30 mg/L
Reported statistics and error estimates:
Not reported
Validity criteria fulfilled:
yes
Conclusions:
Under the test conditions, the EC50 of the test material (containing 99.6 % quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides) for activated sludge after 3 h contact time is 68 mg/L ( 95% CL: 15 and 858 mg/L). The EC10, EC20 and EC80 were determined to be 12, 22 and 215 mg/L, respectively
Executive summary:

An activated sludge respiration inhibition test of the test material (containing 99.6 % quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides) was performed. The toxicity to activated sludge was determined in accordance with OECD Guideline 209, and in compliance with GLP.

This study is assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment it is considered appropriate to read across from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the structurally analogous quaternary dialkyldimethylammonium nitrites.

The test is valid as shown by the EC50 of the reference compound, 3, 5- dichlorophenol (8 mg/L) and the control respiration rates which are within 15 % of each other.

The toxicity of test material to activated sludge was determined at a contact time of 3 h, using various concentrations of the test substance. The inhibitory effect of test material at a particular concentration is expressed as a percentage of the two controls. Under the test conditions, the EC50 of the test material (containing 99.6 % quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides) for activated sludge after 3 h contact time is 68 mg/L (95% CL: 15 and 858 mg/L). The EC10, EC20 and EC80 were determined to be 12, 22 and 215 mg/L, respectively.

Endpoint:
toxicity to microorganisms, other
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1988
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment
Justification for type of information:
Quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment and the counter anion.

In the case of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites that anion will be "nitrite" (NO2−) which will associate with other cations in solution. It is therefore prudent to consider read across from sodium nitrite (CAS Number 7632-00-0, EC Number 231-555-9) in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

This publication is entitled "Toxicity of Inorganic Compounds in the Spirotox Test : A Minaiturized Version of the Spirostomum Ambiguum Test" Nalecz-Jawecki, G & Sawicki, J; Arch. Environ. Contam. Toxicol., 34, 1-5, 1988 as the primary source. In this publication the freshwater ciliata Spirostomum ambiguum was exposed to sodium nitrite solution over 48h in a static test system in order to determine the effects of sodium nitrite to microorganisms acute toxicity and different deformations were recorded.

This publication is also referenced in the Screening Information Dataset (SIDS) Initial Assessment Report for Sodium Nitrite (2005) as a secondary source.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
no guideline followed
Principles of method if other than guideline:
Method: according to Nalecz-Jawecki G & Sawicki J 1988. The freshwater ciliata Spirostomum ambiguum was exposed to sodium nitrite solution over 48h in a static test system in order to determine the effects of sodium nitrite to microorganisms acute toxicity and different deformations were recorded.
GLP compliance:
no
Analytical monitoring:
no
Vehicle:
no
Test organisms (species):
other: Spirostomum ambiguum
Details on inoculum:
- Spirostomum ambiguum is one of the biggest protozoans, 2-3 mm long (Raabe 1970). The strain used in the work was originally collected in Kampinos National Park near Warsaw and has been cultured in the laboratory for more than 20 years.

- Method of cultivation: S. ambiguum was routinely cultured in 5-1 aquariums containing 4 L of natural, unpolluted water originating from a very deep source (pH 5 - 7.5; total hardness 150 mg CaCO3/L). Cultures were maintained at 20–25°C in darkness. Protozoa were fed once a week with a diet of flaked oats and dried alder leaves (50:1). Every 4 weeks, two-thirds of the water in the aquarium was replaced with fresh water.

- Preparation of inoculum for exposure: In order to prepare the protozoa for the test, it was necessary to rinse them from the culturing medium. Several hundred cells were taken from the culture and placed in a 50-ml cylinder. After the cells had fallen to the bottom, water was poured out and organisms were rinsed three times with diluent Diluted Tyrod solution, 1:64 was used; it was made up of 125 mg NaCl, 3.1 mg KCl, 3.1 mg CaCl2, 1.55 mg MgCl2, 15.6 mg NaHCO3, and 0.78 mg NaH2PO4 per liter of deionized water (Milli-Q quality).
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
48 h
Hardness:
2.8 mg/L as CaCO3
Test temperature:
25 °C
pH:
7.4 ± 0.2
Key result
Duration:
24 h
Dose descriptor:
EC50
Effect conc.:
285 mg/L
Conc. based on:
other: NO2-N
Basis for effect:
other: Different deformations
Remarks on result:
other: +/- 135 mg/L
Remarks:
morphological changes such as shortening, bending of the cell, etc.
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
281 mg/L
Conc. based on:
other: NO2-N
Basis for effect:
other: Different deformations
Remarks on result:
other: +/- 137 mg/L
Remarks:
morphological changes such as shortening, bending of the cell, etc.
Key result
Duration:
48 h
Dose descriptor:
EC50
Effect conc.:
421 mg/L
Conc. based on:
test mat.
Basis for effect:
other: Different deformations
Remarks on result:
other: morphological changes such as shortening, bending of the cell, etc.
Validity criteria fulfilled:
not specified
Conclusions:
In a publication entitled "Toxicity of Inorganic Compounds in the Spirotox Test : A Minaiturized Version of the Spirostomum Ambiguum Test" Nalecz-Jawecki, G & Sawicki, J; Arch. Environ. Contam. Toxicol., 34, 1-5, 1988 as the primary source the freshwater ciliata Spirostomum ambiguum was exposed to sodium nitrite solution over 48h in a static test system in order to determine the effects of sodium nitrite to microorganisms acute toxicity and different deformations were recorded.

The 48-hour EC50 and LC50 were 421 and 533 mg/L (expressed as NaNO2), respectively.
Executive summary:

The acute toxicity of the test item (sodium nitrite) to the freshwater ciliata Spirostomum ambiguum was investigated in a published study entitled "Toxicity of Inorganic Compounds in the Spirotox Test : A Minaiturized Version of the Spirostomum Ambiguum Test" Nalecz-Jawecki, G & Sawicki, J; Arch. Environ. Contam. Toxicol., 34, 1-5, 1988 as the primary source.

This publication is also referenced in the Screening Information Dataset (SIDS) Initial Assessment Report for Sodium Nitrite (2005) as a secondary source.

This study is assigned a reliability score of 2 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment and the counter anion, the counter anion in this case being "nitrite" (NO2−), it is considered prudent to read across from sodium nitrite  (CAS Number 7632-00-0, EC Number 231-555-9) in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

In this study the effect of sodium nitrite on deformation (morphological changes such as shortening, bending of the cell, etc.) and mortality of Spirostomun ambiguum (protozoa) was examined. Under the conditions of the study the 48-hour EC50 and LC50 were 421 and 533 mg/L (expressed as NaNO2), respectively.

Endpoint:
activated sludge respiration inhibition testing
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1993
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: Study is conducted according to ISO guideline 8192; nevertheless, the documentation is insufficient for assessment.
Justification for type of information:
Quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment and the counter anion.

In the case of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites that anion will be "nitrite" (NO2−) which will associate with other cations in solution. It is therefore prudent to consider read across from sodium nitrite (CAS Number 7632-00-0, EC Number 231-555-9) in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

In this study the test item was subjected to an activated sludge respiration inhibition test conducted according to ISO guideline 8192.
Reason / purpose for cross-reference:
read-across: supporting information
Qualifier:
according to guideline
Guideline:
ISO 8192 (Water quality - Test for inhibition of oxygen consumption by activated sludge for carbonaceous and ammonium oxidation)
Version / remarks:
The documentation is insufficient for assessment.
GLP compliance:
no
Analytical monitoring:
not specified
Vehicle:
not specified
Test organisms (species):
activated sludge, industrial
Details on inoculum:
- Initial biomass concentration: 1 g/L (dry weight)
Test type:
static
Water media type:
freshwater
Limit test:
no
Total exposure duration:
16 h
pH:
7.0 ±0.2
Nominal and measured concentrations:
Test concentrations (nominal; 30 min): 0, 1, 10, 100 and 1000 mg/LTest concentrations (nominal; 16 h): 0, 1, 100 and 1000 mg/L
Key result
Duration:
30 min
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Nitrite
Basis for effect:
inhibition of total respiration
Key result
Duration:
16 h
Dose descriptor:
EC50
Effect conc.:
> 1 000 mg/L
Nominal / measured:
nominal
Conc. based on:
other: Nitrite
Basis for effect:
inhibition of total respiration
Conclusions:
In an activated sludge respiration inhibition test conducted according to ISO guideline 8192 the EC50 (16 h) of the test item was determined to be > 1000 mg/L.
Executive summary:

The toxicity of the test item to microorganisms was investigate in an activated sludge respiration inhibition test conducted in accordance with ISO guideline 8192. The study was not conducted in accordance with GLP.

This study is assigned a reliability score of 3 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997) since although the study was conducted according to ISO guideline 8192

the documentation is insufficient for assessment.

Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment and the counter anion, the counter anion in this case being "nitrite" (NO2−), it is considered prudent to read across from sodium nitrite  (CAS Number 7632-00-0, EC Number 231-555-9) in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

Under the conditions of the study the EC50 (16 h) of the test item was determined to be > 1000 mg/L.

Description of key information

Read across data is presented in order to demonstrate the toxicity to microorganisms of quaternary ammonium compounds, di-C12 -18 -alkyldimethyl nitrites.

The read across data is taken from Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1 and from sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9. These substances were selected since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations and their counter anions in the aquatic environment.

Furthermore, in certain organic solvents it has been reported that the exchange constants between nitrite and chloride in quaternary ammonium salts (QAS) are approximately equal. [Zhurnal Analiticheskoi Khimii, 2010, Vol. 65, No. 6, pp. 579–584. (E.M. Rakhman’ko, M.S. Markovskaya, L.S. Stanishevskii, Yu.S. Zubenko, A.R. Tsyganov)]

Therefore, it is considered appropriate to read across from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the structurally analogous quaternary dialkyldimethylammonium nitrites. In addition to this the counter anion is "nitrite" (NO2−) which will associate with other cations in solution. It is therefore prudent to consider read across from sodium nitrite in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

Therefore, studies are presented on quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides (1) and on sodium nitrite (3).

Quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides, CAS Number 68391-05-9, EC Number 269-924-1

An activated sludge respiration inhibition test of the test material (containing 99.6 % quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides) was performed. The toxicity to activated sludge was determined in accordance with OECD Guideline 209, and in compliance with GLP.

This study is assigned a reliability score of 1 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment it is considered appropriate to read across from quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides to the structurally analogous quaternary dialkyldimethylammonium nitrites.

The test is valid as shown by the EC50 of the reference compound, 3, 5- dichlorophenol (8 mg/L) and the control respiration rates which are within 15 % of each other.

The toxicity of test material to activated sludge was determined at a contact time of 3 h, using various concentrations of the test substance. The inhibitory effect of test material at a particular concentration is expressed as a percentage of the two controls. Under the test conditions, the EC50 of the test material (containing 99.6 % quaternary ammonium compounds, di-C12-18-alkyldimethyl, chlorides) for activated sludge after 3 h contact time is 68 mg/L (95% CL: 15 and 858 mg/L). The EC10, EC20 and EC80 were determined to be 12, 22 and 215 mg/L, respectively.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9

1.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9

2.

The acute toxicity of the test item (sodium nitrite) to the freshwater ciliata Spirostomum ambiguum was investigated in a published study entitled "Toxicity of Inorganic Compounds in the Spirotox Test : A Minaiturized Version of the Spirostomum Ambiguum Test" Nalecz-Jawecki, G & Sawicki, J; Arch. Environ. Contam. Toxicol., 34, 1-5, 1988 as the primary source.

This publication is also referenced in the Screening Information Dataset (SIDS) Initial Assessment Report for Sodium Nitrite (2005) as a secondary source.

This study is assigned a reliability score of 2 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997).

Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment and the counter anion, the counter anion in this case being "nitrite" (NO2−), it is considered prudent to read across from sodium nitrite  (CAS Number 7632-00-0, EC Number 231-555-9) in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

In this study the effect of sodium nitrite on deformation (morphological changes such as shortening, bending of the cell, etc.) and mortality of Spirostomun ambiguum (protozoa) was examined. Under the conditions of the study the 48-hour EC50 and LC50 were 421 and 533 mg/L (expressed as NaNO2), respectively.

Sodium nitrite, CAS Number 7632-00-0, EC Number 231-555-9

3.

The toxicity of the test item to microorganisms was investigate in an activated sludge respiration inhibition test conducted in accordance with ISO guideline 8192. The study was not conducted in accordance with GLP.

This study is assigned a reliability score of 3 in accordance with the criteria for assessing data quality set forth by Klimisch et al (1997) since although the study was conducted according to ISO guideline 8192

the documentation is insufficient for assessment.

Since quaternary ammonium substances are to be expected to dissociate into their quaternary ammonium cations in the aquatic environment and the counter anion, the counter anion in this case being "nitrite" (NO2−), it is considered prudent to read across from sodium nitrite  (CAS Number 7632-00-0, EC Number 231-555-9) in order to realise a fuller impression of the environmental properties of Quaternary ammonium compounds, di-C12-18-alkyldimethyl, nitrites.

Under the conditions of the study the EC50 (16 h) of the test item was determined to be > 1000 mg/L.

Key value for chemical safety assessment

EC50 for microorganisms:
68 mg/L
EC10 or NOEC for microorganisms:
12 mg/L

Additional information

In addition to the information stated above the Screening Information Datset (SIDS) Initial Assessment Report for Sodium NItrite (2005) also references an additional study on the toxicity to microorganisms of sodium nitrite :-

Ostrensky and Lemos (1993) reported a 96h EC50 (mobility) of 7886 mg NaNO2/L and 96h NOEC (mobility) of 3740 NaNO2/L for Tetraselmis chuii. (Ostrensky A and Lemos D (1993). Effects of Ammonium and Nitrite on Growth of the Marine Microalgae Tetraselmis chuii. Actas IV Congreso Nac. Acuicult. 485-489).

Categories Display

EU Privacy Disclaimer

This website uses cookies to ensure you get the best experience on our websites.