Flue dust, portland cement

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-04-07 to 2010-04-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, according to the OECD 209 technical guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

yes

Principles of method if other than guideline:

In contrast to the guideline, it was decided to use copper (II) suphate pentahydrate instead of the prescribed 3,5-dichlorophenol. The usage of dichlorophenol is unsuitable for routine lab use.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item was weighed out directly in an Erlenmeyer flask and fille dup with 250 mL autoclaved tap water. Afterwards the solution was stirred for 24 h.
- Eluate: Synthetic waste water acc. to OECD 209
- Differential loading: 100 - 180 - 320 - 580 - 1000 mg/L without adjustment of pH
+ 580 - 1000 mg/L (adjusted to pH7 with HCl c=1mol/L)
- Controls: 2 control samples without test item were included, one at the beginning and one at the end of the test.

Test organisms (species):

activated sludge

Details on inoculum:

- Preparation of inoculum for exposure: Non adapted activated sludge from the sewage plant at sarstedt (Municipal sewage treatment plant of 31157 Sarstedt, Germany)
- Pretreatment: Washed twice with autoclaved tap water and adjusted to a dry weight concentration of 4.0 g/L +/- 10 %
- Dry sludge concentration: 4.24 g/L

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Hardness:

Not determined

Test temperature:

20.8 °C

pH:

6.71

Salinity:

Not determined

Details on test conditions:

TEST SYSTEM
- Test vessel: 500 mL Erlenmeyer flask, DIN 12380
- Aeration: Permanent
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2
- Composition of test medium: 200 mL inoculum + 16 mL synthetic waste water (acc. to the guideline) + 284 mL autoclaved tap water + test substance concentration (directly weighing of 500 mg)



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Determination of respiration rates subsequently to an incubation period of 3 h

TEST CONCENTRATIONS

- Range finding study
Inhibitions in the Preliminary Tests
Inhibitions in the Preliminary Tests
Nominal Test Item
Concentration [mg/L] Inhibition
[%]
10 (pH: 7.23) 2
100 (pH: 7.38) 2
1000 (pH: 9.70) 75

Reference substance (positive control):

yes

Remarks:

Copper (II) sulphate pentahydrate p.a.

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

743 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: (660-830)

Details on results:

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: EC50: 100 mg/L

Reported statistics and error estimates:

The EC-values of the test item (without adjustment of pH) were calculated by sigmoidal dose-response regression using software GraphPad Prism4, GRAPHPAD SOFTWARE, INC. Calculation of the confidence intervals for the EC50-value was carried out using standard procedures.

The EC50-value of the reference item was calculated by probit analysis using software SIGMAPLOT (Windows), SPSS CORPORATION. Probit values were taken from WEBER (1986).

From the oxygen depletion the respiration rates were calculated in mgO₂/(L×h).

The inhibition of the respiration rates from the test and reference item concentrations in % was calculated in comparison with the respiration rates from the control.

Respiration Rates and Inhibition of Respiration of Test and Reference Item Concentrations

	Concentration	Respiration rate	Inhibition
	[mg/L]	[mgO2/(L×h)]	[%]
Test item		1000		4.4		87
		580		30.0		12
		320		36.8		-8
		180		39.6		-16
		100		30.0		12
		1000 ^#		36.8		-8
		580 ^#		36.0		-6
Control 1		–		33.6		–
Control 2		–		34.4		–
Reference item		180		9.2		73
		100		17.2		49
		58		24.0		29

EC-Values with Confidence Interval of Flue Dust S

	EC-values	Confidence interval
	[mg/L]	P = 95 [%] [mg/L]
EC20	615	-
EC50	743	660-830
EC80	923	-

Raw Data and Respiration Rates of Test Item, Control and Reference Item

	Concentration	Oxygen-concentration*	Measured distance**	Respiration rate
	[mg/L]	[mgO2/L]	[mm/3 min]	[mgO2/(L×h)]
Test item	1000	9.21	11	4.4
	580	7.76	75	30.0
	320	7.84	92	36.8
	180	7.24	99	39.6
	100	5.99	75	30.0
	1000 ^#	7.11	92	36.8
	580 ^#	6.34	90	36.0
Control 1	–	5.20	84	33.6
Control 2	–	6.87	86	34.4
Reference item	180	8.41	23	9.2
	100	6.91	43	17.2
	58	6.92	60	24.0

Validity criteria fulfilled:

yes

Conclusions:

The test item Flue Dust S is not toxic at concentrations to activated sludge of a municipal sewage treatment plant.

Executive summary:

A Respiration Inhibition Test according to OECD Guideline No. 209 was performed with the test item FlueDust S from 2010-04-07 to 2010-04-08 at Dr.U.Noack-Laboratorien, D-31157 Sarstedt. Test system was activated sludge of the municipal treatment plant of D-31157 Sarstedt. The test was performed under static conditions with the nominal concentrations 100 - 180 - 320 - 580 - 1000 mg/L without adjustment of pH and 580 - 1000 mg/L adjusted to pH 7 to differentiate between pH effects and test item effects. The respiration rates of control, reference and test item concentrations were measured after a contact time of three hours, and the inhibitory effects of the test and reference item were determined in comparison to the control respiration rates. The concentration effect relationship of the test item concentrations without adjustment of pH is shown in Figure 1. The inhibition of the test item ranged from -16 % to 87 %. The neutralised test item concentrations showed inhibitions from  -8 % to -6 %. Therefore it can be concluded that the determined inhibition is due to the low pH value of the test solution but no effect caused by the test item.

EC-Values with Confidence Interval of Flue Dust S

	EC-values	Confidence interval
	[mg/L]	P = 95 [%] [mg/L]
EC20	615	-
EC50	743	660-830
EC80	923	-

In order to check the activity of the test system and the test conditions a reference test was carried out with copper (II) sulphate pentahydrate as reference item and the reference toxicity was determined. The EC₅₀-value for the reference item was 100 mg/L.

Validity criteria of the test guideline were fulfilled.

The test item Flue Dust S is not toxic at concentrations </= 615 mg/L to activated sludge of a municipal sewage treatment plant.

Reference 2

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-04-21 to 2010-04-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study, according to the OECD 209 technical guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

yes

Principles of method if other than guideline:

In contrast to the guideline, it was decided to use copper (II) suphate pentahydrate instead of the prescribed 3,5-dichlorophenol. The usage of dichlorophenol is unsuitable for routine lab use.

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The test item was weighed out directly in an Erlenmeyer flask and filled up with 250 mL autoclaved tap water. Afterwards the solution was stirred for 24 h.
- Eluate: Synthetic waste water acc. to OECD 209
- Differential loading: 100 - 180 - 320 - 580 - 1000 mg/L without adjustment of pH
+ 580 - 1000 mg/L (adjusted to pH7 with HCl c=1mol/L)
- Controls: 2 control samples without test item were included, one at the beginning and one at the end of the test.

Test organisms (species):

activated sludge

Details on inoculum:

- Preparation of inoculum for exposure: Non adapted activated sludge from the sewage plant at sarstedt (Municipal sewage treatment plant of 31157 Sarstedt, Germany)
- Pretreatment: Washed twice with autoclaved tap water and adjusted to a dry weight concentration of 4.0 g/L +/- 10 %
- Dry sludge concentration: 4.22 g/L

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Hardness:

Not determined

Test temperature:

20.8 °C

pH:

6.90

Salinity:

Not determined

Details on test conditions:

TEST SYSTEM
- Test vessel: 500 mL Erlenmeyer flask, DIN 12380
- Aeration: Permanent
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 2
- Composition of test medium: 200 mL inoculum + 16 mL synthetic waste water (acc. to the guideline) + 284 mL autoclaved tap water + test substance concentration (directly weighing of 500 mg)



EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Determination of respiration rates subsequently to an incubation period of 3 h

TEST CONCENTRATIONS

- Range finding study
Inhibitions in the Preliminary Tests
Nominal Test Item
Concentration [mg/L] Inhibition
[%]
10 (pH: 7.31) -7
100 (pH: 7.46) -9
1000 (pH: 10.09) 81

Reference substance (positive control):

yes

Remarks:

Copper (II) sulphate pentahydrate p.a.

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

596 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: (533-669)

Details on results:

- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: None

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- Relevant effect levels: EC50: 106 mg/L

Reported statistics and error estimates:

The EC-values of the test item (without adjustment of pH) were calculated by sigmoidal dose-response regression using software GraphPad Prism4, GRAPHPAD SOFTWARE, INC. Calculation of the confidence intervals for the EC50-value was carried out using standard procedures.

The EC50-value of the reference item was calculated by probit analysis using software SIGMAPLOT (Windows), SPSS CORPORATION. Probit values were taken from WEBER (1986).

From the oxygen depletion the respiration rates were calculated in mgO₂/(L×h).

The inhibition of the respiration rates from the test and reference item concentrations in % was calculated in comparison with the respiration rates from the control.

Respiration Rates and Inhibition of Respiration of Test and Reference Item Concentrations

	Concentration	Respiration rate	Inhibition *
	[mg/L]	[mgO2/(L×h)]	[%]
Test item		1000		1.6		94
		580		14.0		46
		320		27.2		-5
		180		24.0		8
		100		26.8		-3
		1000 ^#		28.4		-9
		580 ^#		28.0		-8
Control 1		–		26.4		–
Control 2		–		25.6		–
Reference item		180		7.6		71
		100		13.6		48
		58		19.6		25

EC-Values with Confidence Interval of Flue Dust T

	EC-values	Confidence interval
	[mg/L]	P = 95 [%] [mg/L]
EC20	489	-
EC50	596	533-669
EC80	728	-

Raw Data and Respiration Rates of Test Item, Control and Reference Item

	Concentration	Oxygen-concentration*	Measured distance**	Respiration rate
	[mg/L]	[mgO2/L]	[mm/3 min]	[mgO2/(L×h)]
Test item	1000	8.67	4	1.6
	580	7.41	35	14.0
	320	6.84	68	27.2
	180	5.56	60	24.0
	100	6.65	67	26.8
	1000 ^#	6.11	71	28.4
	580 ^#	6.72	70	28.0
Control 1	–	5.86	66	26.4
Control 2	–	6.95	64	25.6
Reference item	180	8.23	19	7.6
	100	7.98	34	13.6
	58	6.39	49	19.6

Validity criteria fulfilled:

yes

Conclusions:

The test item Flue Dust T is not toxic at concentrations to activated sludge of a municipal sewage treatment plant.

Executive summary:

A Respiration Inhibition Test according to OECD Guideline No. 209 was performed with the test item Flue Dust T (batch number: 12-2009) from 2010-04-21 to 2010-04-22 at Dr.U.Noack-Laboratorien, D-31157 Sarstedt. Test system was activated sludge of the municipal treatment plant of D-31157 Sarstedt. The test was performed under static conditions with the nominal concentrations 100 - 180 - 320 - 580 - 1000 mg/L without adjustment of pH and 580 - 1000 mg/L adjusted to pH 7 to differentiate between pH effects and test item effects. The respiration rates of control, reference and test item concentrations were measured after a contact time of three hours, and the inhibitory effects of the test and reference item were determined in comparison to the control respiration rates. The concentration effect relationship of the test item concentrations without adjustment of pH is shown in Figure 1. The inhibition of the test item ranged from -5 % to 94 %. The neutralised test item concentrations showed inhibitions ≤ -8 %. Therefore it can be concluded that the determined inhibition is due to the low pH value of the test solution but no effect caused by the test item.

EC-Values with Confidence Interval of Flue Dust T

	EC-values	Confidence interval
	[mg/L]	P = 95 [%] [mg/L]
EC20	489	-
EC50	596	533-669
EC80	728	-

In order to check the activity of the test system and the test conditions a reference test was carried out with copper (II) sulphate pentahydrate as reference item and the reference toxicity was determined. The EC₅₀-value for the reference item was 106 mg/L.

Validity criteria of the test guideline were fulfilled.

The test item Flue Dust T is not toxic at concentrations </= 489 mg/L to activated sludge of a municipal sewage treatment plant.

Description of key information

No acute toxicity to aquatic algae due to a Klimisch 1 study result.

Key value for chemical safety assessment

EC50 for microorganisms:

596 mg/L

EC10 or NOEC for microorganisms:

425 mg/L

Additional information

Short-term toxicity testing to microorganisms was carried out due to GLP and thus resulting in a Klimish 1 score.

Two Respiration Inhibition Tests according to OECD Guideline No. 209 were performed with the two test items Flue Dust S and Flue Dust T (batch number: 12-2009) from 2010-04-21 to 2010-04-22 at DR.U.NoAcK- LABORATORIEN, D-31157 Sarstedt. Test system was activated sludge of the municipal treatment plant of D-31157 Sarstedt. The test was performed under static conditions with the nominal concentrations 100 - 180 - 320 - 580 - 1000 mg/L without adjustment of pH and 580 - 1000 mg/L adjusted to pH 7 to differentiate between pH effects and test item effects. The respiration rates of control, reference and test item concentrations were measured after a contact time of three hours, and the inhibitory effects of the test and reference item were determined in comparison to the control respiration rates. The inhibition of the test item ranged from -5 % to 94 % for Flue Dust T and from -8 to 87 % for Flue Dust S. The neutralised test item concentrations showed inhibitions < -8 %. Therefore it can be concluded that the determined inhibition is due to the low pH value of the test solution but no effect caused by the test item.

The test item Flue Dust T is not toxic at concentrations < 489 mg/L to activated sludge of a municipal sewage treatment plant.

The test item Flue Dust S is not toxic at concentrations < 615 mg/L to activated sludge of a municipal sewage treatment plant.