Amines, C12-14-tert-alkyl, mixed sec-Bu and iso-Bu phosphates

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 Jul - 10 Aug 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

no analytical determination was performed

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Hessisches Ministerium für Umwelt, Energie, Landwirtschaft und Verbraucherschutz

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: a water accommondated fraction was performed: for each test concentration the required aomunt of test item was dispersed in dilution water and was shaked at 130 rpm for 24 h at room temperature; subsequently the suspensions were filtered through glass fibre filters and eluates were used for the test
- Differential loading: 0.5, 1.0, 2.0, 4.0 and 8.0 mg/L
- Controls: yes

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: STRAUS (clone 5)
- Source: obtained from BASF SE, Limburgerhof, Germany; the strain originated from the Institut National de Recherche Chimique Appliquee, France in 1978 and cultivated since then at BASF using medium M4 acc. Elendt
- Feeding during test: no feeding during the test
- Food type: for cultivation: with Desmodesmus subspicatus- and/or Chlorella sp.cells

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES: daphnids are transferred into fresh medium one day before the study start in order to ensure the lest animals were less than 24 hours old.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Test temperature:

20.8 -21.1 °C (control)
20.7 - 21.2 °C (test item concentration)

pH:

8.02 - 8.70 (control)
8.03 - 8.92 (test item concentration)

Dissolved oxygen:

8.3 - 8.5 mg O2/L (control)
7.7 - 8.4 mg O2/L (test item concentration)

Nominal and measured concentrations:

control, 0.5, 1.0, 2.0, 4.0, 8.0 loading of the test item (mg/L)

Details on test conditions:

TEST SYSTEM
Test vessel:
- Type: open
- Material, size, headspace, fill volume: 50 ml-glass beakers, 50 ml volume
- No. of organisms per vessel: 5 animals
- No. of vessels per concentration (replicates): 20
- No. of vessels per control (replicates): 20

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: test item was diluted with test water which was prepared according to OECD Guidance Document on Aquatic Toxicity Testing of Difficult Substances and Mixtures, 2000
- Culture medium different from test medium: no
- Intervals of water quality measurement: pH, temperature and oxygen content was measured at test start and test end

OTHER TEST CONDITIONS
- Photoperiod: light/dark cycle 16h of light and 8 hours of darkness

EFFECT PARAMETERS MEASURED: immobility at 24 and 48 h

VEHICLE CONTROL PERFORMED: no

RANGE-FINDING STUDY
- Test concentrations: 5.1, 10.8, and 100 mg/L
- Results used to determine the conditions for the definitive study: Based on the results of the screening /range-finding test (20% inhibition at 5 mg/L and 90% inhibition at 10 mg/L) the concentrations of the main test was performed

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EL10

Effect conc.:

0.56 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Remarks on result:

other: 95% confidentail limits

Remarks:

0.01 - 0.94 mg/L

Duration:

48 h

Dose descriptor:

other: EL20

Effect conc.:

0.72 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Remarks on result:

other: 95% confidential limits

Remarks:

0.05 - 1.15 mg/L

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

1.13 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Remarks on result:

other: 95% confidential limits

Remarks:

0.44 - 2.54 mg/L

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

0.5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Duration:

48 h

Dose descriptor:

LOELR

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Details on results:

- Mortality of control: no mortality was observed
- Others: Biological Results are summarized in the tables 1-3 within the section: "Any other information on results incl tables"

Results with reference substance (positive control):

- ECx: EL50 (24 h) 1.0 mg/L

Reported statistics and error estimates:

The inhibition values as well as the statistical calculation were done according to the guideline using the commercial computer program ToxRat Version 2.10.

Validity criteria were fulfilled:

In the control without any test item no dead animals were  observed.

In the test solutions the oxygen concentration did not drop below a value of 60% of the saturation value (5 mg O2/L).

Analytical Determination:

The EC50-value was calculated on the basis of the nominal concentrations tested as being agreed with the sponsor because there is no real analytical method for the active substance which could identify the test item (UVCB substance) in the aqueous phase at very low concentrations (0.5 - 10 mg/L). Only HPLC would detect this UVCB-substance but at concentrations which are 3 or more magnitudes higher. Due to these methodical problems the sponsor decided to waive the analytical part and to choose the nominal concentrations.

Table 1: Screening Test (Range-finder)

Loading of the lest item (mg/L) (Eluates/”WAF”)	Ration immobilized Daphnia magna [%]
	t 24 h	t 48 h
Control (0)	0 [0/10]	0 [0/10]
5	10 [10/10]	20 [2/10]
10	70 [7/10]	9 [9/10]
100	100 [10/10]	100 [10/10]

 

Table 2: Biological Results Definitive test

Loading of the lest item (mg/L)	Ratio immobilised Daphnia magna [%]
(Eluates/”WAF”)	24 h	48 h
Control (0)	0 [0/20]1	0 [0/20] 1
0.5	0 [0/20]1	0 [0/20]1
1.0	40 [8/20]1	60 [12/20]1
2.0	60 [12/20]1	75 [15/20]1
4.0	65 [13/20]1	100 [20/20]1
8.0	65 [13/20]1	100 [20/20]1

¹⁾  Numbers in brackets give the absolute numbers of immobilised animals at eachconcentration

Table 3: Endpoints

On the Basis of the nominal Concentrations [ma EC306-227-4 /,,Additin RC 3740")/L]
		t24h	t48h
	EL10	0.81	n.d.
95% -CL	lower	n.d.	n.d.
	upper	n.d.	n.d.
	EL20	1.15	0.59
95% -CL	lower	n.d.	n.d.
	upper	n.d.	n.d.
	EL50	2.27	1.15
95% -CL	lower	n.d.	n.d.
	upper	n.d.	n.d.
Mortality	LOEL	2.00	:s: 1.00
	NOEL  1.00	< 1.00

n.d.       not determined due to mathematical reasons (inappropriate data)

Validity criteria fulfilled:

yes

Remarks:

see detailed information in section "Any other information on results incl. tables"

Description of key information

EL50 (48h) = 1.13 mg/L (OECD 202, Daphnia magna, nominal)

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

1.13 mg/L

Additional information

One study is available, in which the short term toxicity to invertebrates was investigated according to GLP and OECD guideline 202 (SGS Institut Fresenius GmbH, 2012) using the test organism Daphnia magna. The test was performed under static test conditions and with the test item concentrations of 0.5, 1.0, 2.0, 4.0, 8.0 mg/L (WAF, nominal loading rate). Since no analytical method was applicable the endpoints were evaluated using the nominal concentrations. The EL50 (48 h) was therefore determined to be at 1.13 mg/L based on the nominal concentrations.