1,3-bis(2-methylphenyl)guanidinium m-[[p-anilinophenyl]azo]benzenesulfonate

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:

From 2014-08-05 to 2014-08-14

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

yes

Remarks:

(since the test item is poorly water soluble, the standard method for the preparation of aqueous media was modified. The test was performed with a saturated solution of the test item)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.2 (Acute Toxicity for Daphnia)

GLP compliance:

no

Test material

Test material information

1

Reference substance name:

1-3(Diphenylguanidinium) m-[[p-anilinophenyl]azo]benzenesulfonate

EC Number:

941-151-0

Molecular formula:

C31H28N6SO3

IUPAC Name:

1-3(Diphenylguanidinium) m-[[p-anilinophenyl]azo]benzenesulfonate

Test material form:

solid: particulate/powder

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method of preparation: Saturation of the experimental solution:
100 mg of test item was dispersed in 1 liters test medium
Stirring for 24 hours.
Filtration through a PTFE of 0.22 μm to remove undissolved test item
Obtention of a 100% v/v saturated solution.
Series of dilution to give further test concentration for the range finding study and the main test.

- Application: daphnies were placed in test tube containing 10 mL of the saturated solution test

- Controls: control group maintained under identical conditions but not exposed to the test item
- Vehicle: none
- Evidence of undissolved material (e.g. precipitate, surface film, etc): no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Source: in-house subcontracting laboratory cultures
- Age at study initiation: less than 24 hours old
- Method of breeding:
Breeding medium: Adult Daphnia were maintained in a synthetic medium as defined by the standard NF EN ISO 6341
Temperature: 18°C
Continuous lightning (intensity of 1000 lux on the containers surface)
No aeration
Containers: 3 litre glass beaker
Breeding charge: around 50 spawners per glass beaker
Breeding medium renewal frequency: twice per week
Spawners renewal frequency: around every 3 weeks
Test medium acclimatation of the spawners: no period of acclimatation

- Feeding during test: no

ACCLIMATION
- Acclimation period: no

QUARANTINE (wild caught)
- Duration:no quarantine

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

225 mg/L +/- 50 mg/L as CaCO3

Test temperature:

18°C to 22°C

Nominal and measured concentrations:

Nominal test concentration, in % v/v saturated solution :
a - control
b- 18 % v/v
c- 27 % v/v
d- 40 % v/v
e- 60 % v/v
f- 100 % v/v

Details on test conditions:

TEST SYSTEM
- Test vessel: glass test tube of 10 mL
- Type : closed
- Material, size, headspace, fill volume: 10 mL glass test tube
- Aeration: no
- Renewal rate of test solution: no renewal
- No. of organisms per vessel: 5 organisms
- No. of vessels per concentration (replicates): 4 replicates
- No. of vessels per control (replicates): 4 replicates

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: medium prepared according to the standard NF EN ISO 6341 (see table 2 for composition).

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: continous lighting
- Light intensity: no light

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- immobilisation of the daphnies

TEST CONCENTRATIONS
- Spacing factor for test concentrations: mean of √2.4
- Range finding study: yes
- Test concentrations: 18, 27, 40, 60 and 100 % v/v saturated solution
- Results used to determine the conditions for the definitive study: cumulative immobilization data from the range-finding study

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr207)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC0

Effect conc.:

ca. 27 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: (95% CL: 84.4 - > 100 mg/l)

Effect concentrations 3

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

- Results with reference substance valid? yes. EC50 - 24 hr = 1.1 mg/L (2014/08/11)

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Under the experimental conditions:
* 48h - EC50 > 100 mg/L
* 48h - NOEC = 27 mg/L
* 48h - LOEC > 100 mg/L

Executive summary:

Introduction

A study was performed to assess the acute toxicity of the test item to Daphnia magna.The method followed was designed to be compatible with OECD Guidelines. for Testing of Chemicals (April 2004) No 202,"Daphnia sp., Acute Immobilisation Test", referenced as method C.2 of Council Regulation (EC) No. 440/2008.

 

Methods

 

Following a preliminary range-finding test, twenty daphnids (4 replicates of 5 animals) were exposed to an aqueous solution of the test item at concentrations of 1.0, 3.5, 10, 35 and 100% v/v saturated solution for 48 hours at a temperature of 20°C to 21°C under static test conditions.

The test item solutions were prepared by stirring an excess of test item in test medium using a propeller stirrer at approximately 1500 rpm for 24 hours. After the stirring period any undissolved test item was removed by filtration (0.2 µm Gelman Acrocap filter) to produce a 100% v/v saturated solution of the test item. This saturated solution was then further diluted as necessary, to provide the remaining test groups at 18, 27, 40, 60 and 100 mg/L. The number of immobilized Daphnia were recorded after 24 and 48 hours.

 

Results

 

Exposure of Daphnia magna to the test item gave the following results :

-         EC50 – 48 hr > 100 mg/L (IC_95%84.4 - > 100)

-         NOEC = 27 mg/l

-         LOEC > 100 mg/l