3-(isotridecyloxy)propylamine

Toxicity to microorganisms

Administrative data

Endpoint:

activated sludge respiration inhibition testing

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012-02-29 to 2012-03-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study, GLP, complete report and good identification of test substance

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2012

Report date:

2012

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 209 (Activated Sludge, Respiration Inhibition Test

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-(isodecyloxy)propylamine

EC Number:

250-056-7

EC Name:

3-(isodecyloxy)propylamine

Molecular formula:

C13H29NO

IUPAC Name:

3-(isodecyloxy)propylamine

Test material form:

other: Liquid

Details on test material:

- Name of test material (as cited in study report): 3-(Isodecyloxy)propylamine
- CAS no 30113-45-2
- Physical state: Liquid, light yellow
- Analytical purity: 95.3 %
- Purity test date: 2012-07-03
- Lot/batch No.: 890000022898
- Expiration date of the lot/batch: 2019-09-22
- Stability under test conditions: Not applicable
- Storage condition of test material: Room temperature, protected from light, in the original container, stored under N2

Sampling and analysis

Analytical monitoring:

no

Test solutions

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, filled up with 200 mL demieralised water and stirred overnight.
- Eluate: Test medium:
250 mL inoculum
16 mL synthetic waste water
234 mL autoclaved tap water
- Differential loading: 6.56 - 13.1 - 26.2 - 52.5 - 104.9 mg/L (test item)
- Controls: Test medium without test or reference item.

Test organisms

Test organisms (species):

activated sludge

Details on inoculum:

- Preparation of inoculum for exposure: Non adapted activated sludge from the sewage plant at Hildesheim (Municipal sewage treatment plant of 31137 Hildesheim, Germany)
- Pretreatment: Washed twice with autoclaved tap water and adjusted to a dry weight concentration of 3 g/L +/- 10 %. The sludge was used within 24 h after stirring.
- Dry sludge concentration: 2.73 g/L corresponding to 1.40 g/L suspended solids in the test vessels

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

3 h

Test conditions

Hardness:

Not determined

Test temperature:

20.6 °C

pH:

7.35 (activated sludge)
7.87 (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 Item Concentration Pretreatment pH Inhibition [%]
[mg/L]
10 Stirring overnight 8.22 0
100 8.74 89
1000 10.79 95
1000 10.26 97
1000 7.85* 93

Reference substance (positive control):

yes

Remarks:

Copper (II) sulphate pentahydrate p.a.

Results and discussion

Effect concentrations

Duration:

3 h

Dose descriptor:

EC10

Effect conc.:

13 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

inhibition of total respiration

Remarks:

respiration rate

Remarks on result:

other: (11.6-14.6)

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 NOEC was determined by calculation of statistical significance of the inhibition of respiration in comparison to the control. One Way Analysis of Variance (ANOVA) and DUNNETT’s test was used for NOEC calculations. When running a One Way Analysis of Variance a Normality test and an Equal Variance test was done first. P-values for both Normality and Equal Variance test are 0.05. The a-value (acceptable probability of incorrectly concluding that there is a difference) is a=0.05.

The EC-values of the test item were calculated by sigmoidal dose-response regression and the EC50-value of the reference item by linear regression using software GraphPad Prism, GRAPHPAD SOFTWARE, INC. Calculation of the confidence intervals for the EC50-values were carried out using standard procedures.

Any other information on results incl. tables

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	35.2	25.1	–	–
	2	36.4	26.0
	3	37.2	26.6
	4	32.0	22.9
	5	32.0	22.9
	6	31.2	22.3
58	1	24.8		27	27
	2	24.4		28
	3	25.6		25
100	1	17.2		49	49
	2	17.2		49
	3	17.2		49
180	1	7.6		78	76
	2	8.0		76
	3	8.4		75

Oxygen Uptake 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 [%]
6.25	1	32.0	6	0
	2	33.6	1
	3	33.6	1
	4	36.8	-8
	5	33.2	2
12.5	1	30.0	12	8
	2	33.6	1
	3	28.8	15
	4	32.4	5
	5	32.0	6
25	1	18.0	47	56
	2	13.6	60
	3	13.2	61
	4	14.4	58
	5	16.4	52
50	1	5.6	84	84
	2	5.2	85
	3	6.0	82
	4	6.0	82
	5	4.8	86
100	1	3.2	91	91
	2	2.8	92
	3	3.2	91
	4	2.8	92
	5	3.2	91

EC-Values with Confidence Interval of 3-(Isodecyloxy)propylamine

	EC-values	Confidence interval
	[mg a.i./L]	P = 95 [%] [mg a.i./L]
NOEC	12.5	–
EC10	13.0	11.6 – 14.6
EC20	16.0	14.6 – 17.4
EC50	23.6	22.4 – 24.8
EC80	38.7	34.7 – 43.7

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The NOEC of the test item 3-(Isodecyloxy)propylamine is 12.5 mg/L. The EC10 and EC50 are resp. 13 and 23.6 mg/L
The test item is not toxic at concentrations

Executive summary:

A Respiration Inhibition Test with activated sludge according to OECD Guideline No. 209 was carried out with the test item 3-(Isodecyloxy)propylamine (batch number: 890000022898) from 2012-02-29 to 2012-03-15, with the experimental phase on 2012-03-15 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 6.25 – 12.5 – 25.0 – 50.0 – 100 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 0 % to 91 % .

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.

EC-Values with Confidence Interval of 3-(Isodecyloxy)propylamine

		EC-values		Confidence interval
		[mg a.i./L]		P = 95 [%] [mg a.i./L]
NOEC		12.5		–
EC10		13.0		11.6 – 14.6
EC20		16.0		14.6 – 17.4
EC50		23.6		22.4 – 24.8
EC80		38.7		34.7 – 43.7

                                              

The EC10 of the test item 3 -(Isodecyloxy)propylamine is 13 mg a.i./L. The test item is not toxic at concentrations </= 12.5 mg a.i./L to activated sludge of a municipal sewage treatment plant.