Octanoic acid, compound with octylamine (1:1)

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-05-11 to 2016-05-13

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Octanoic acid, compound with octylamine (1:1)

EC Number:

241-477-7

EC Name:

Octanoic acid, compound with octylamine (1:1)

Cas Number:

17463-34-2

Molecular formula:

C8H19N.C8H16O2

IUPAC Name:

octan-1-amine; octanoic acid

Test material form:

other: Pale brown, waxy solid

Details on test material:

Test Item: Caprylamine caprylate

Chemical name: Octanoic acid, compound with octylamine

CAS number: 17463-34-2

Molecular formula: C8H19N.C8H16O2

Molecular weight: 273.5 g/mol

Appearance: Pale brown, waxy solid

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Determination of the test item monitoring
All concentration levels and the control were analytically verified via
LC-MS/MS at the start (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours).
The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000). Details of the analytical method are presented.

Sampling for the analytical
At the start of the exposure intervals (0 and 24 hours), sampling was
carried out from the newly prepared test concentrations.
At the end of the exposure intervals (24 and 48 hours), samples were taken directly from the test vessels.

Range (target)
Recoveries of the test item should be within ± 20% of the nominal or initially measured concentrations.

Test solutions

Vehicle:

no

Details on test solutions:

Stock solution
The stock solution (10.0 mg/L of the test item were weighed out) was freshly prepared with dilution water (Table 2) before the start of the exposure intervals (at 0 and 24 hours).

Treatment
The stock solution was mixed thoroughly by manual agitation.

Test concentrations
5 test item concentrations in a geometric series with a separation factor of 2, prepared by dilution of the stock solution with dilution water, were tested as follows:
0.625 - 1.25 - 2.50 - 5.00 - 10.0 mg/L
The test concentrations mentioned above were selected based on the results of a non-GLP preliminary range finding test.

Control
Dilution water without test item incubated under the same conditions as the test groups.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test system
Daphnia magna STRAUS (Clone 5).

Reason for the selection of the test system
Daphnia magna is the preferred species in accordance with the test guideline and is bred at the test facility.

Origin
Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), 14195 Berlin, Germany

Breeder
Noack Laboratorien GmbH, Käthe-Paulus-Str. 1, 31157 Sarstedt, Germany

Culture
In glass vessels (2 - 3 L capacity) with approximately 1.8 L culture medium, at 20 ± 2 °C, in an incubator, 16 hours illumination, light intensity of max. 20 µE.m-2 . s-1 (max. 1340 lx)

Culture medium
Elendt M4, according to OECD 202, Annex 3 (2004), modified to a total hardness of 160 to 180 mg CaCO3/L, is used. The composition of the culture medium is presented

Culture feeding
The culture daphnids were fed at least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella subcapitata and Desmodesmus subspicatus, with an algae cell density of > 106 cells/mL. The algae were cultured at the test facility.

Origin of the food algae
Sammlung von Algenkulturen (SAG),
Pflanzenphysiologisches Institut der Universität Göttingen, Nikolausberger Weg 18, 37073 Göttingen, Germany

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test conditions

Hardness:

Dilution water
2016-05-11: 174 mg CaCO3/L
2016-05-12: 178 mg CaCO3/L

Test temperature:

Dilution water
2016-05-11: 20.2 °C
2016-05-12: 20.1 °C

pH:

Dilution water
2016-05-11: 8.53
2016-05-12: 8.77

Dissolved oxygen:

Dilution water
2016-05-11: 8.89 mg/L
2016-05-12: 8.77 mg/L

Conductivity:

Dilution water
2016-05-11: 473 µS/cm
2016-05-12: 484 µS/cm

Nominal and measured concentrations:

The concentrations of the test item Caprylamine caprylate were analytically verified via LC-MS/MS at the start of the exposure intervals (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours) in all tested concentration levels and in the control. Details of the analytical method are presented.
The measured concentrations of the test item Caprylamine caprylate were in the range of 95 to 116% of the nominal values at the start of the exposure intervals (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours). The analytical results are presented.
The measured test item concentrations remained stable within ± 20% of the nominal concentrations throughout the exposure period. This indicates that the test item concentrations were successfully maintained for the duration of the test.

Details on test conditions:

Test duration
48 hours

Test vessels
Glass beakers (4 (ID) x 7 (H) cm), 50 mL capacity, loosely covered with watch glasses

Renewal of the test solutions
The test solutions were renewed after 24 hours. For this purpose, a second set of test vessels was filled with the freshly prepared test solutions and the daphnids were transferred by pipette (see ‘Application’).

Dilution water
Same as culture medium (see Table 2)

Number of daphnids and replicates
20 daphnids, divided into 4 replicates, each with 5 daphnids, were used per concentration level and control.

Age of the daphnids at the start of the exposure
Less than 24 hours old daphnids from a healthy stock were used for the study. Juvenile daphnids were removed from the culture vessels at the latest 24 hours before the start of the exposure and discarded. The juveniles born within the following period of max. 24 hours preceding the exposure were used for the test. No first brood progeny was used for the test.

Acclimatisation
Acclimatisation of the daphnids was not necessary, because the dilution water was equivalent to the culture medium.

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC10

Effect conc.:

3.09 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC10

Effect conc.:

1.57 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

6.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

5.42 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 6

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

The percentage of immobility for the reference item was determined after 24 hours. The EC50-value with 95% confidence limits (CI) was calculated by sigmoidal dose-response regression. The EC50-value for the most recent of the monthly performed reference tests was:
EC50: 2.08 mg/L (CI 1.00 - 4.00 mg/L)
The EC50-value of the reference item potassium dichromate after 24 hours is within the required concentration range of 0.6 - 2.4 mg/L of quality criteria according to AQS P 9/2 (02/2000) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Any other information on results incl. tables

Measured Concentrations of the Test ItemCaprylamine caprylateduring the Definitive Test

Sampling date	2016-05-11 Start of the  exposure interval 0 hours		2016-05-12 End of the  exposure interval 24 hours		2016-05-12 Start of the  exposure interval 24 hours		2016-05-13 End of the  exposure interval 48 hours
Start of analysis	2016-05-11		2016-05-12		2016-05-12		2016-05-13
Nominal test item concentration [mg/L]	Caprylamine caprylate
	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
10.0	11.6	116	11.4	114	11.4	114	10.5	105
5.00	5.28	106	5.40	108	5.38	108	5.02	100
2.50	2.53	101	2.68	107	2.70	108	2.38	95
1.25	1.34	107	1.44	115	1.38	110	1.25	100
0.625	0.678	108	0.714	114	0.678	108	0.599	96
Control	< LOQ		< LOQ		< LOQ		< LOQ

Meas. conc.= measured concentration of the test item, mean value of 2 injections, dilution factors taken into account

%                  = percent of the nominal concentrations of the test item

LOQ            = limit of quantification (20 µg/L of the test item)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the nominal concentrations of the test item Caprylamine caprylate, the 48 hour EC50 for Daphnia magna was 5.42 mg/L
(95% confidence limits: 4.37 – 6.82 mg/L).

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the test item Caprylamine caprylate were determined according to OECD 202 (2004) from 2016-05-11 to 2016-05-13at the test facility.

The study was conducted under semi-static conditions over a period of 48 hours with five nominal concentrations of the test item Caprylamine caprylateranging from 0.625 to 10.0 mg/Lin a geometric series with a separation factor of 2.

The test item Caprylamine caprylateis a pale brown, waxy solid with a water solubility of 2.0 - 3.5 g/L (20 °C, cmc).A stock solution with a nominal concentration of 10.0 mg/L of the test item was freshly prepared with dilution water before the start of the exposure intervals (at 0 and 24 hours), mixed thoroughly by manual agitation and was used as the highest test concentration and as a stock solution for the preparation of the further concentration levels.

All test item concentrations were visually clear throughout the exposure period.

Twenty daphnids (divided into 4 replicates with 5 daphnids each) were exposed to each concentration level and the control.

The concentrations of the test item Caprylamine caprylate were analytically verified via LC-MS/MS at the start of the exposure intervals (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours) in all tested concentration levels and in the control. Details of the analytical method are presented.

The measured concentrations of the test itemCaprylamine caprylate were in the range of 95 to 116% of the nominal values at the start of the exposure intervals (0 and 24 hours) and at the end of the exposure intervals (24 and 48 hours).The analytical results are presented.

The measured test item concentrations remained stable within ± 20% of the nominal concentrations throughout the exposure period. This indicates that the test item concentrations were successfully maintained for the duration of the test. Therefore, the effect valuesare based on the nominal concentrations of the test item Caprylamine caprylate.

The validity criteria of the test guideline were fulfilled.

 

  EC₁₀-, EC₅₀- (with 95% Confidence Limits) and EC₁₀₀-Values

(based on the nominal concentrations of the test item)

Caprylaminecaprylate
Effect levels	Test duration   [hours]	Toxicity endpoint based on the nominal concentrations the test item [mg/L]
EC10 (with 95% confidence limits)	24	3.09	(Cl: 2.36 – 3.91)
	48	1.57	(Cl: 0.955 – 2.26)
EC50 (with 95% confidence limits)	24	6.50	(CI: 5.72 – 7.43)
	48	5.42	(Cl: 4.37 – 6.82)
EC100	24	> 10.0
	48	> 10.0