Tosylchloramide sodium

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to microorganisms, other

Remarks:

A test with Chloramine-T trihydrate was performed according to NEN 6512 2nd design and ISO TC 147 SC 5 WG 1 N 52.

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods with acceptable restrictions

Remarks:

little detail is given in the final report. The information presented is however sufficient for evaluation of the effects on sludge bacteria.

Qualifier:

according to guideline

Guideline:

other: NEN 6512 (Dutch national guideline)

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: ISO TC 147 SC 5 WG 1 N 52

Deviations:

not specified

Principles of method if other than guideline:

A test with Chloramine-T trihydrate was performed according to NEN 6512 2nd design and ISO TC 147 SC 5 WG 1 N 52. Three different tests were performed:
Respiration inhibition of activated sludge from a domestic water treatment plant (Duiven, The Netherlands) with sodium acetate as a substrate.
Respiration inhibition of nitrifying sludge from an enrichment culture using ammonium carbonate as nutrition and addition.
Inhibition of methane production in anaerobic methane fermentation sludge.

GLP compliance:

no

Remarks:

pre-GLP

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

- Strain: activated sludge 
- Source/supplier: WWTP, Nieuwgraaf, Arnhem, The Netherlands
- Laboratory culture: no
- Method of cultivation: no
- Pretreatment: no pretreatment. 
- Controls: yes
- Initial cell concentration: not described 

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

5 min

Nominal and measured concentrations:

- Concentrations: 2, 10, 50 and 500 mg/l

Details on test conditions:

- Type: aquatic
- Exposure vessel type: glass
- Number of replicates: single-fold

Reference substance (positive control):

no

Duration:

5 min

Dose descriptor:

EC50

Effect conc.:

ca. 5 mg/L

Nominal / measured:

nominal

Basis for effect:

other: respiration inhibition activated sludge

Duration:

5 min

Dose descriptor:

EC50

Effect conc.:

700 mg/L

Nominal / measured:

nominal

Basis for effect:

other: respiration inhibition nitrifying sludge

Duration:

5 min

Dose descriptor:

EC50

Effect conc.:

1 000 mg/L

Nominal / measured:

nominal

Basis for effect:

other: methane production inhibition sludge

Reported statistics and error estimates:

ECn values determined by linear interpolation of data.

Respiration inhibition activated sludge
Concentration Chloramine-T trihydrate mg/L in sludge	% Respiration inhibition
500	100
50	80
10	70
2	0-20

Respiration inhibition nitrifying sludge
Concentration Chloramine-T trihydrate mg/L in sludge	% Respiration inhibition
2500	90
1000	81
500	15
100	10
50	6

Methane production inhibition sludge After 40 days incubation with Chloramine-T trihydrate , average DOC of Chloramine-T trihydrate is 0.3 mg/mg
Sample	DOC values	Average
Blanc	200* 126	126
glucose	126 120 138	128
Glucose + 1 mg Chloramine-T trihydrate/L	135 113	124
Glucose + 10 mg Chloramine-T trihydrate/L	130 120	125
Glucose + 100 mg Chloramine-T trihydrate/L	161 159	160
Glucose + 1000 mg Chloramine-T trihydrate/L	478 467	473
Glucose + 10000 mg Chloramine-T trihydrate/L	2497 2697	2584

*this value was disturbed by suspended sludge

There is a great difference in sensitivity between the respiration inhibition of sludge and nitrifying sludge. The methane production from glucose is slowed down significantly at 100-1000 mg/L at 10000 mg/L this is totally inhibited.  

Validity criteria fulfilled:

not specified

Conclusions:

Respiration inhibition activated sludge: EC50 5 mg/L
Respiration inhibition nitrifying sludge: EC50 700 mg/L
Methane production inhibition sludge: EC50 1000 mg/L

Executive summary:

A test with Tosylchloramide sodium, trihydrate was performed according to NEN 6512 2^nd design and ISO TC 147 SC 5 WG 1 N 52. Three different tests were performed: Respiration inhibition of activated sludge from a domestic water treatment plant (Duiven, The Netherlands) with sodium acetate as a substrate.    

Respiration inhibition of nitrifying sludge from an enrichment culture using ammonium carbonate as nutrition and addition.

Inhibition of methane production in anaerobic methane fermentation sludge.  

There is a great difference in sensitivity between the respiration inhibition of sludge and nitrifying sludge. The methane production from glucose is slowed down significantly at 100-1000 mg/L at 10000 mg/L this is totally inhibited.  

Reference 2

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1988

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Not enough data available to assess validity.

Qualifier:

according to guideline

Guideline:

other: Robra test (German national guideline)

Principles of method if other than guideline:

Inhibition of respiration of Pseudomonas putida was determined after exposure of an active culture with Halamid for several minutes. 

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Test organisms (species):

Pseudomonas putida

Test type:

static

Water media type:

freshwater

Limit test:

no

Details on test conditions:

Type: aquatic

Key result

Duration:

30 min

Dose descriptor:

EC10

Effect conc.:

ca. 0.001 other: %

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Validity criteria fulfilled:

not applicable

Conclusions:

Tosylchloramide sodium, trihydrate inhibits bacterial respiration at low concentrations (0.001%) with almost 100% inhibition at 0.5% Tosylchloramide sodium, trihydrate.

Reference 3

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP study according to protocol.

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

not required

Vehicle:

no

Details on test solutions:

Chloramine T was added to the incubation vessels using a stock solution of approximately 1.0 g/L. The stock solutionwas prepared by dissolving 0.10 g Chloramine T in 100 mL deionized water.

Test organisms (species):

activated sludge of a predominantly domestic sewage

Details on inoculum:

Unadapted secondary activated sludge was obtained (05-12-2011) from the WWTP Nieuwgraaf in Duiven. The WWTP Duiven is an activated sludge plant treating predominantly domestic wastewater. 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. The dry weight of the activated sludge in the incubation vessels was 1.5 g/L.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test temperature:

19.9 - 20.9 °C

pH:

6.9 - 7.5

Nominal and measured concentrations:

Nominal: 10, 20, 40, 80, 160 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 300 mL Erlenmeyer flasks
- Aeration: no
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2

TEST MEDIUM / WATER PARAMETERS
Synthetic sewage feed and stocks:
The synthetic sewage feed was made by dissolving the following amounts of substances in 1 liter of deionized water: 16 g peptone, 11 g meat extract, 3 g urea, 0.7 g NaCl, 0.3 g CaCl2, 0.2 g MgSO4.7H2O, 2.8 g K2HPO4. In the incubation vessels 1.6 mL of the synthetic sewage feed is added in a total volume of 50 mL.

OTHER TEST CONDITIONS
- Adjustment of pH: No

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : repsiration rate

TEST CONCENTRATIONS
- Spacing factor for test concentrations: 2
- Range finding study: Not performed

Reference substance (positive control):

yes

Remarks:

3,5-dichlorophenol

Key result

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

37 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: 95% C.I. 16-78 mg/L

Key result

Duration:

3 h

Dose descriptor:

EC10

Effect conc.:

14 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Results with reference substance (positive control):

EC50 of the reference compound is 12 mg/L on the first day and 15 mg/L on the second day, which is within the prescribed range of 2 to 25 mg/L.

Reported statistics and error estimates:

Probit analysis.

Validity criteria fulfilled:

yes

Conclusions:

The test is performed accordding to standard guideline under GLP, therefore the test can be considered reliable. Validity criteria were met, therefore the test is valid.

Executive summary:

In order to predict effects of chemicals in the environment and in biological waste water treatment plants, an activated sludge respiration inhibition test was performed. The toxicity to activated sludge was determined in accordance with OECD Test Guideline 209, and in compliance with the OECD principles of Good Laboratory Practice. The test was performed on two subsequent days after collection of the activated sludge. The test is valid as shown by the coefficient of variation of the replicates of the control oxygen uptake rates (5% on the first day and 6% on the second day), the EC50 of the reference compound 3,5-dichlorophenol (12 mg/L on the first day and 15 mg/L on the second day) and the average control oxygen uptake rate of 18 mg O2/g dry weight/hour on the first day and 36 mg O2/g dry weight/hour on the second day. The oxygen uptake rate on the first day is slightly lower than prescribed. Accurate oxygen uptake rate measurements were very well possible as shown by the very low coefficient of variation. Therefore this is a minor deviation which does not invalidate the test. The increase in the oxygen uptake rates on the second day compared to the first day does not significantly influence the sensitivity of the activated sludge as shown by the EC50 values of the reference substance. The toxicity of Halamid PG to activated sludge was determined at a contact time of 3 hours, using various concentrations of the test substance. The inhibitory effect of Halamid PG at a particular concentration is expressed as a percentage of the two controls. From the results EC values were calculated. The EC50 of Halamid PG for activated sludge after 3 hours contact time is 37 mg/L with 95% confidence limits of 16 and 78 mg/L. The EC10, EC20 and EC80 are 14, 20 and 71 mg/L, respectively. Halamid PG is therefore considered harmful to activated sludge.

Reference 4

Endpoint:

toxicity to microorganisms

Adequacy of study:

other information

Reliability:

2 (reliable with restrictions)

Principles of method if other than guideline:

Method: other: Bioassay for monitoring biochemical methane potential and anaerobic toxicity, W.F. Owen et al, Water Research, 13 (1979).pp. 485-492

GLP compliance:

no

Analytical monitoring:

yes

Test organisms (species):

anaerobic bacteria from a domestic water treatment plant

Details on test conditions:

Type: aquatic

Duration:

40 d

Dose descriptor:

EC50

Effect conc.:

ca. 1 000 mg/L

Duration:

40 d

Dose descriptor:

EC100

Effect conc.:

ca. 10 000 mg/L

RS-Freetext:
RESULTS: EXPOSED
- Nominal/measured concentrations: nominal 
- Effect data/Element values: EC 50 and 100
RESULTS CONTROL: no effect
STATISTICAL RESULTS: ECn values determined by linear
interpolation of data.

Reference 5

Endpoint:

toxicity to microorganisms

Adequacy of study:

other information

Reliability:

2 (reliable with restrictions)

Principles of method if other than guideline:

Method: other: Blok, J. Respirometric measurments on activated sludge, Water Research Vol. 8 (1974), pp.11-18

GLP compliance:

no

Analytical monitoring:

yes

Test organisms (species):

other: other bacteria: Nitrifying bacteria

Details on test conditions:

Type: aquatic

Dose descriptor:

EC50

Effect conc.:

ca. 700 mg/L

RS-Freetext:
RESULTS: EXPOSED
- Nominal/measured concentrations: nominal 
- Effect data/Element values: EC 50 
RESULTS CONTROL: no effect
STATISTICAL RESULTS: ECn values determined by linear
interpolation of data.

Description of key information

Multiple guideline studies regarding toxicity to microorganisms are available. The study with the lowest EC50 value is considered key.

A test with Tosylchloramide sodium, trihydrate was performed according to NEN 6512 2nd design and ISO TC 147 SC5WG1N52. Three different tests were performed: Respiration inhibition of activated sludge from a domestic water treatment plant (Duiven, The Netherlands) with sodium acetate as a substrate. Respiration inhibition of nitrifying sludge from an enrichment culture using ammonium carbonate as nutrition and addition.Inhibition of methane production in anaerobic methane fermentation sludge. There is a great difference in sensitivity between the respiration inhibition of sludge and nitrifying sludge. The methane production from glucose was slowed down significantly at 100-1000 mg/L, and totally inhibited at 10000 mg/L. The EC50 of respiration inhibition of activated, nitrifying and methane production inhibition sludge was 5, 700 and 1000 mg/L respectively.

Key value for chemical safety assessment

EC50 for microorganisms:

5 mg/L

Additional information

The key value deriving the PNEC STP microorganism is based on the most conservative value derived from a klimisch 2 study by Blok (1982). Additional key and supporting studies by Bessems (1988) and Geerts (2012) are available.