Fatty acid chlorides, C8-14 (even numbered), reaction products with glycine

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-01-10 to 2012-02-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

not specified

GLP compliance:

yes

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: No vehicle, directly weighing of the stock solution (10 g a.i./L)
- Eluate: Synthetic waste water acc. to OECD 209
- Differential loading: 1000 - 580 - 320 - 180 - 100 mg/L
- Controls: Test medium without test or reference substance. 6 replicates: Triplicates at the beginning of the test + triplicates 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 3 g/L +/- 10 %
- Dry sludge concentration: 3.05 g/L

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Hardness:

Not determined

Test temperature:

20-21 °C

pH:

7.88 (activated sludge)
7.37 (synthetic waste water)

Salinity:

None, freshwater

Details on test conditions:

TEST SYSTEM
- Test vessel: 1000 mL Erlenmeyer flask, DIN 12380
- Aeration: Permanent
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 3
- Composition of test medium:
250 mL inoculum
16 mL synthetic waste water
234 mL autoclaved tap water
Test or reference item concentration (directly weighed into each test vessel)


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
After 3 h incubation, the oxygen concentration of each control. reference item and test item replicate was determined by means of oximeter immediately after transferring the solution to the measuring cell and the oxygen depletion was recorded for at least 3 minutes on a flat bed recorder.

TEST CONCENTRATIONS

- Range finding study
Inhibition of Respiration (Preliminary Test)
Nominal Test
Concentration
[mg a.i./L] Inhibition[%]
10 2
100 -15
1000 89

Reference substance (positive control):

yes

Remarks:

Copper (II) sulphate pentahydrate p.a.

Duration:

3 h

Dose descriptor:

NOEC

Effect conc.:

320 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Duration:

3 h

Dose descriptor:

EC10

Effect conc.:

372 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: 347-398 (95% CI)

Duration:

3 h

Dose descriptor:

EC50

Effect conc.:

791 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: 733-847 (95% CI)

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: 129 mg/L

 EC-Values and NOEC ofHostapon SG (a.i.)

	Effect values (nominal a.i.)	Confidence interval
	[mg/L]	P = 95 [%] [mg/L]
NOEC	320	–
EC10	372	347 – 398
EC20	452	422 – 486
EC50	791	733 – 847
EC80	> 1000	n.d.

Oxygen Uptake Rates, Specific Respiration Rates and Inhibition of the Respiration of the Control and Reference Item Concentrations

Reference Item Concentration [mg/L]	Repl.	Oxygen Uptake Rate R [mg O2/L×h]	Specific Respiration Rate Rs [mg O2/g×h]	Inhibition [%]	Mean Inhibition [%]
Control	1	33.6	22.0	–	–
	2	34.0	22.3
	3	33.2	21.8
	4	36.0	23.6
	5	34.4	22.6
	6	33.6	22.0
58	1	29.2		14	13
	2	31.2		9
	3	28.4		17
100	1	21.6		37	38
	2	21.6		37
	3	20.0		41
180	1	12.4		64	65
	2	12.0		65
	3	11.6		66

Oxygen Uptake Rates, Specific Respiration Rates and Inhibition of the Respiration of the Test Item Concentrations

Test item Concentration [mg a.i./L]	Repl.	Oxygen Uptake Rate R [mg O2/L×h]	Inhibition [%]	Mean Inhibition [%]
100	1	42.0	-23	-33
	2	45.6	-34
	3	48.0	-41
	4	44.0	-29
	5	46.8	-37
180	1	43.2	-27	-22
	2	40.8	-20
	3	41.6	-22
	4	43.6	-28
	5	38.0	-11
320	1	32.8	4	5
	2	32.4	5
	3	33.6	1
	4	32.0	6
	5	31.2	9
580	1	22.8	33	32
	2	23.6	31
	3	22.4	34
	4	22.8	33
	5	24.0	30
1000	1	13.6	60	62
	2	13.6	60
	3	12.8	62
	4	12.4	64
	5	12.0	65

Validity criteria fulfilled:

yes

Conclusions:

The NOEC of the active ingredient is 320 mg/L, the EC10 is 372 mg/L and the EC50 791 mg/L to activated sludge of a municipal sewage treatment plant.

Executive summary:

A Respiration Inhibition Test with activated sludge according to OECD Guideline No. 209 was carried out with the test item Hostapon SG (batch number: ESD 0006364) from 2012-01-10 to 2012-02-07, with the experimental phase on 2012-02-07 at Dr.U.Noack-Laboratorien, D-31157 Sarstedt. Test system was activated sludge of the municipal treatment plant of D-31137 Hildesheim. The test was carried out under static conditions with the nominal active ingredient concentrations 100 - 180 - 320 - 580 - 1000 mg/L. The respiration rates of the control, reference and test item replicates 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 mean inhibition of respiration for the test item replicates ranged from -33 % to 62 %.

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 129 mg/L.

Table 1:   EC-Values and NOEC of Hostapon SG (a.i.)

	Effect values (nominal a.i.)	Confidence interval
	[mg/L]	P = 95 [%] [mg/L]
NOEC	320	–
EC10	372	347 – 398
EC20	452	422 – 486
EC50	791	733 – 847
EC80	> 1000	n.d.

Description of key information

The NOEC of the active ingredient is 320 mg/L, the EC10 is 372 mg/L and the EC50 791 mg/L to activated sludge of a municipal sewage treatment plant.

Key value for chemical safety assessment

EC50 for microorganisms:

791 mg/L