Sodium 3-(4-methyl-2-nitrophenoxy)propanesulphonate

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:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

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:

Sodium 3-(4-methyl-2-nitrophenoxy)propanesulphonate

EC Number:

279-282-4

EC Name:

Sodium 3-(4-methyl-2-nitrophenoxy)propanesulphonate

Cas Number:

79817-52-0

Molecular formula:

C10H13NO6S.Na

IUPAC Name:

sodium 3-(4-methyl-2-nitrophenoxy)propane-1-sulfonate

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

The limit concentration and the control were analytically verified via HPLC-DAD at the start (0 hours) and at the end of the exposure (48 hours). The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000).

Test solutions

Vehicle:

no

Details on test solutions:

Limit concentration
A limit concentration of 100 mg/L of the test item was tested. The limit concentration was based on the results of a non-GLP preliminary range finding test under static conditions.

Preparation of the test concentration
The limit concentration (100 mg/L test item were weighed out) was freshly prepared with dilution water prior to the start of the exposure (at 0 hours). The limit concentration was mixed thoroughly by manual agitation until the test item was completely dissolved.

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 system
Daphnia magna STRAUS (Clone 5)

Reason for the selectionof 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. 1500 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.

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:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Total hardness [mg CaCO3/L] : 171

Test temperature:

19.8 °C

pH:

Water Quality Parameters at the Start of the Exposure (0 hours)
(measured in one additional replicate of the limit concentration and the control)
Nominal
test item concentration
[mg/L] pH-value DissolvedO2 concentration [mg/L]
100 7.46 9.31
Control 7.50 9.54


Water Quality Parameters at the End of the Exposure (48 hours)
(measured in all replicates of the limit concentration and the control)
Nominal
test item concentration[mg/L] pH-values Dissolved O2 concentration [mg/L]
Replicates Replicates
1 2 3 4 1 2 3 4
100 7.25 7.27 7.34 7.37 8.46 7.34 8.21 8.34
Control 7.19 7.15 7.17 7.22 8.48 8.39 8.38 8.30

Dissolved oxygen:

see above

Conductivity:

2017-03-07 [µS/cm] : 420

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 in each test replicate (see ‘Application’)

Dilution water
Same as culture medium

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

Age of the daphnids
Less than 24 hours old daphnids from a healthy stock were used for at the start of the exposure 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.

Acclimatization
Acclimatization 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. 1500 lx

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

Feeding
The daphnids were not fed during the study.

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

other: EC 10/50/100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

other: EC 10/50/100

Effect conc.:

> 100 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 potassium dichromate (SIGMA-ALDRICH, batch number MKBV0900V, purity 99.0%, CAS RN 7778-50-9) was determined after 24 hours from 2017-03-09 to 2017-03-10.

EC50-Value (with 95% confidence limits) of the Reference Item Potassium dichromate
based on nominal concentrations mg/L, (0 - 24 hours)
Current Study Valid Range
EC50 2.05 mg/L
0.6 - 2.4 mg/L, acc. to AQS P 9/2 (02/2000); clone 5
0.6 - 2.1 mg/L, acc. to OECD 202 (2004); clone A
95% confidence limits 2.00 - 3.71 mg/L

Reported statistics and error estimates:

Methods of evaluation
The effects of the limit concentration were deduced directly from the observed immobilization rates. Since the measured test item concentrations were within ± 20% of the nominal concentration, all given effect concentrations (EC10 / 50 / 100) were based on the nominal concentration of the test item Nitrophenoxypropansulfonsäure, Na-Salz

EC-values and statistical analyses for reference item
An EC50-value was calculated for the reference item by sigmoidal dose-response regression. The respective 95% confidence limits were 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

    Immobilization Rates [%] after 24 and 48 hours of Exposure in the Definitive Test

                (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal test item concentration [mg/L]	IMMOBILIZATION [%]
	24 hours					48 hours
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

 

 

 Absolute Numbers of immobile Daphnids after 24 and 48 hours of Exposure in the Definitive Test

                (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal test item concentration [mg/L]	Number of immobile Daphnids / Total number of Daphnids
	24 hours					48 hours
	Replicates					Replicates
	1	2	3	4	MV	1	2	3	4	MV
100	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20
Control	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20	0 / 5	0 / 5	0 / 5	0 / 5	0 / 20

 

The effect concentrations, based on the nominal concentration of the test itemNitrophenoxypropansulfonsäure, Na-Salz, are presented inTable1.

 

 

 

Measured Exposure Concentrations during the Definitive Test

 

The concentration of the test item Nitrophenoxypropansulfonsäure, Na-Salzwas analytically verified via HPLC-DAD at the start of the exposure (0 hours) and at the end of the exposure (48 hours) in the limit concentration and in the control.

The measured concentration in the limit concentration was 96% of the nominal concentration at the start of the exposure (0 hours) and 99% of the nominal concentration at the end of the exposure (48 hours).

The measured test item concentrations were within +/- 20% of the nominal concentration. This indicates that the test item concentration was successfully maintained for the duration of the test.

 

   Measured Concentrations of the Test Itemduring the Definitive Test

Sampling date:	2017-03-07 Start of the exposure, 0 hours		2017-03-09 End of the exposure, 48 hours
Start of analysis:	2017-03-07		2017-03-09
Nominal test item concentration [mg/L]	Nitrophenoxypropansulfonsäure, Na-Salz
	Meas. conc. [mg/L]	%	Meas. conc. [mg/L]	%
100	96.0	96	99.0	99
Control	< LOQ		< LOQ

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

%                     = percent of the nominal concentration of the test item

LOQ                = limit of quantification of the analytical method (10.0 mg test item/L)

 

 

  Water Quality Parameters

 

The measured water quality parameters(i.e. pH-value, dissolved oxygen concentration, total water hardness and water temperature) were within the acceptable limits. During the test period, the temperature in the incubator was 19 - 20 °C.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In the limit concentration 100 mg/L of the test item Nitrophenoxypropansulfonsäure, Na-Salz no effects on Daphnia magna were observed.

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS), the effects of the limit concentration 100 mg/L of the test item Nitrophenoxypropansulfonsäure, Na-Salz (batch number: DAFA015079) were determined at the test facility according to OECD 202 (2004) from 2017-03-07 to 2017-03-09.

The limit test was conducted under static conditions over a period of 48 hours.The limit concentration was slightly yellowish and clearly dissolved during the exposure period.Twenty daphnids, divided into 4 replicates with 5 daphnids each, were exposed to the limit concentration and the control.

The concentration of the test item Nitrophenoxypropansulfonsäure, Na-Salz was analytically verified via HPLC-DAD at the start of the exposure (0 hours) and at the end of the exposure (48 hours) in the limit concentration and in the control.

The measured concentration in the limit concentration was 96% of the nominal concentration at the start of the exposure (0 hours) and 99% of the nominal concentration at the end of the exposure (48 hours).

The measured test item concentrations were within ± 20% of the nominal concentration. This indicates that the test item concentration was successfully maintained for the duration of the test. Therefore, the given effect concentrations are based on the nominal concentration of the test item Nitrophenoxypropansulfonsäure, Na-Salz.

The validity criteria of the test guideline were fulfilled.

 

EC_{10 / 50 / 100}-Values (based on the nominal concentration of the test item)

Effect Concentrations	Test duration [h]	Nitrophenoxypropansulfonsäure, Na-Salz Nominal test item concentration [mg/L]
EC10 / 50 / 100	24	> 100
	48	> 100