Reaction mass of methyl N-octadecylterephthalamate and 37437-26-6

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:

2016-04-25 to 2016-04-28

Reliability:

1 (reliable without restriction)

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

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Methyl N-octadecylterephthala​mate and N,​N'-​Dioctadecylterephtha​lamide

IUPAC Name:

Reaction mass of Methyl N-octadecylterephthala​mate and N,​N'-​Dioctadecylterephtha​lamide

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Sampling and analysis

Analytical monitoring:

no

Test solutions

Vehicle:

no

Details on test solutions:

Limit loading level
A WAF with a loading level of 100 mg/L was tested in a limit test.
Per definition of the WAF, all terms related to the concentration level were given as loading level because partly dissolved compounds and mixtures cannot be related to concentrations.

Water Accommodated Fraction (WAF)
In view of the difficulties associated with the evaluation of aquatic toxicity of poorly water soluble test items, a modification of the standard method for the preparation of aqueous media was performed. An approach endorsed by several important regulatory authorities in the EU and elsewhere (ECETOC 1996 and OECD 2000) is to expose the organisms to a Water Accommodated Fraction (WAF) of the test item in cases where the test item is a complex mixture and is poorly soluble in water and in the permitted solvents. Using this approach, aqueous media was prepared by mixing the test item with water for a prolonged period sufficient to ensure equilibration between the test item and the water phase. At the completion of mixing and following a settlement period, the test item phase was separated by siphon and the test organisms exposed to the aqueous phase, the WAF (which may contain dissolved and/or suspended and/or emulsified fractions of the test item mixture). Exposure was expressed in terms of the original concentration of the test item in water at the start of the mixing period (loading rate) irrespective of the actual concentration of the test item in the WAF.

Preparation of the water accommodated fraction
the water accommodated fraction (WAF) was prepared with a loading level of 100 mg/L as specified below.
100 mg of the test item were weighed out and transferred into a brown glass flask with 1 L of dilution water (Table 2). This dispersion was shaken with a rotating shaker for 24 hours with 20 rpm at room temperature. After a separation phase of 1 hour, the aqueous phase or WAF was removed by siphon from the approximate centre of the water body to receive the 100 mg/L loading level.

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

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Test species
Daphnia magna STRAUS (Clone 5).

Reason for the selection of the test species
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

Culture medium
Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L, is used. The composition of the culture medium is presented in Table 2.
 

Composition of the Culture Medium according to ELENDT (1990)
Component Concentration [mg/L]
CaCl2 x 2 H2O 147
MgSO4 x 7 H2O 123
KCl 5.80
NaHCO3 64.8
Na2SiO3 4.30
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18
Na2EDTA x 2 H2O 5.00
FeSO4 x 7 H2O 1.99
H3BO3 0.29
MnCl2 x 4 H2O 0.36
LiCl 0.30
SrCl2 x 6 H2O 0.15
RbCl 0.071
NaBr 0.016
Na2MoO4 x 2 H2O 0.063
CuCl x 2 H2O 0.017
ZnCl2 0.013
CoCl2 x 6 H2O 0.010
KJ 0.00325
Na2SeO3 0.00219
NH4VO3 0.000575
Thiaminhydrochloride 0.075
Cyanocobalamine 0.0010
Biotin 0.00075
pH 8.2  0.8

Culture feeding
The culture daphnids are 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 are 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:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Dilution water dated:
0 hours: 2016-04-26 Total hardness [mg CaCO3/L]: 178

Test temperature:

18 - 22°C, constant within ± 1°C
2016-04-26 : 20.3 °C

pH:

Nominal loading level
of the test item
[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
100 (WAF) 7.58 7.57 7.57 7.58 7.20 7.13 7.40 7.57
Control 7.30 7.49 7.53 7.60 7.79 7.88 8.02 7.94

Dissolved oxygen:

Nominal loading level
of the test item
[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
100 (WAF) 7.58 7.57 7.57 7.58 7.20 7.13 7.40 7.57
Control 7.30 7.49 7.53 7.60 7.79 7.88 8.02 7.94

Nominal and measured concentrations:

Nominal: 100 mg/l

Details on test conditions:

Test method The study was performed with a static test design.

Test duration
48 hours

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

Test volume
20 mL

Dilution water Same as culture medium (see Table )

Number of daphnids
20 daphnids, divided into 4 replicates, each with 5 daphnids, were and replicates used for the WAF and the control.

Age of the daphnids at the start of the exposure
Less than 24 hours old daphnidsat the start of the exposure 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.

Application
20 g test solution per replicate was weighed out into each test vessel. This corresponds to 20 mL per test vessel. The daphnids were inserted with a small amount of dilution water by a pipette.

Test temperature (target)
18 - 22 °C, constant within ± 1 °C

Illumination (target)
Diffuse light, light intensity of max. 20 µEm-2  s-1 (max. 1340 lx)

Photoperiod (target)
16/8 hours light/dark cycle

Feeding
The daphnids were not fed during the study.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EL10

Effect conc.:

> 100 other:

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EL10

Effect conc.:

> 100 other:

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 other:

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 other:

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 5

Duration:

24 h

Dose descriptor:

EL100

Effect conc.:

> 100 other:

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Effect concentrations 6

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

> 100 other:

Nominal / measured:

not specified

Conc. based on:

test mat.

Basis for effect:

other: immobilisation

Results with reference substance (positive control):

The percentage of immobility for the reference item was determined after 24 hours.

Table 5: EC50-Value of the Reference Item Potassium dichromate after 24 hours of exposure
based on nominal concentrations mg/L

Current study: Valid range
2016-04-05 to 2016-04-06
Immobilisation after 24 hours
EC50 1.95 mg/L 0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000)
1.14 - 3.30 mg/L 0.6 - 2.1 mg/L, acc. to OECD 202 (2004)

95 % confidence limits 1.14 - 3.30 mg/L

Reported statistics and error estimates:

Methods of evaluation
The effects of the WAF were deduced directly from the observed immobilisation rates. The effect levels (EL10 / 50 / 100) were based on the nominal loading
level of 100 mg/L of the test item. Per definition of the WAF, all terms related to concentrations have to be given as loading levels because partly dissolved
compounds and mixtures cannot be related to concentrations.

EC-values and statistical analyses for reference item
An EC50-value was calculated only for the reference item by
sigmoidal dose-response regression. The respective 95 % confidence limits for the EC50 was calculated from the standard error and the t-distribution.
All calculations were carried out from the best-fit values with the software GraphPad Prism5.

Software
All data were computer-processed and rounded for presentation. Consequently, minor variations may occur from the original figures if manual calculations based on the original figures are made subsequently. Calculations were made using the following software:
- GraphPad Prism5, GRAPHPAD SOFTWARE, INC.
- Excel, MICROSOFT CORPORATION

Any other information on results incl. tables

Water Quality Parameters at the Start of the Exposure (0 hours)

                     (measured in one additional replicate (without daphnids) of the WAF and the control)

Nominal loading level of the test item [mg/L]	pH-value	Dissolved O2concentration [mg/L]
100 (WAF)	7.81	7.64
Control	8.83	9.95

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the Water Accommodated Fraction with a nominal loading of 100 mg/L of the test item HOSTAGEL HT 300, no effects on Daphnia magna were
observed.

Executive summary:

    Summary

 

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of a Water Accommodated Fraction (WAF) with a nominal loading of 100 mg/L of the test itemHOSTAGELHT 300(batch number:DEA14F316) were determined according to OECD 202 (2004) from2016-04‑25 to 2016-04-28, with the definitive exposure phase from 2016‑04-26 to 2016-04-28, at the test facility.

An approach endorsed by several important regulatory authorities in the EU and elsewhere (ECETOC 1996 and OECD 2000), is to expose the organisms to a Water Accommodated Fraction (WAF) of the test item in cases where the test item is a complex mixture and is poorly soluble in water and in the permitted solvents.

Using this approach, the aqueous medium was prepared by shaking the test item with dilution water at20 rpmfor 24 hoursat room temperatureto ensure equilibration between the test item and the water phase. 100 mgof the test item were weighed out and transferred into a brown glass flask filled with 1000 mL of dilution water(seeTable2).After completion of shaking and following a 1‑hour settlement period, the test item phase was separated by siphon from the approximate center of the water body to receive the WAFwith a nominal loading of 100 mg/L.The WAF was clear throughout the exposure period.

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

No analytical monitoring of the test item was carried out due to the sponsor’s request.

The effect levels based on the nominal loading of test itemHOSTAGELHT 300are given.

The validity criteria of the test guideline were fulfilled.

 

 

 

In the Water Accommodated Fraction with a nominal loading of 100 mg/L of the
test itemHOSTAGEL HT 300, no effects onDaphnia magnawere observed.

 

 

 

Table1:   EL₁₀-, EL₅₀-and EL₁₀₀-Values

(based on the nominal loading of the test itemHOSTAGELHT 300)

Effect levels	Test duration [hours]	Toxicity endpoint based on the nominal loading of the test item
		[mg/L]
EL10 / 50 / 100	24	> 100
	48	> 100