4-[(4-chloro-2-nitrophenyl)azo]-3-hydroxy-N-(2-methylphenyl)naphthalene-2-carboxamide

Toxicity to terrestrial plants

Administrative data

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

2009-07-31 to 2009-08-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study meets requirements of OECD Guideline 211 and GLP requirements with no deviations.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

3-hydroxy-N-(o-tolyl)-4-[(2,4,5-trichlorophenyl)azo]naphthalene-2-carboxamide

EC Number:

229-440-3

EC Name:

3-hydroxy-N-(o-tolyl)-4-[(2,4,5-trichlorophenyl)azo]naphthalene-2-carboxamide

Cas Number:

6535-46-2

IUPAC Name:

3-hydroxy-N-(2-methylphenyl)-4-[(2,4,5-trichlorophenyl)diazenyl]-2-naphthamide

Sampling and analysis

Analytical monitoring:

no

Details on sampling:

No analytical monitoring

Test substrate

Vehicle:

yes

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): Calculated amounts of the test item were mixed thoroughly with quartz sand in a glass vessel (1 % of the entire soil amount per concentration). The treated quartz sand was mixed carefully with the soil with a mixer to insure a homogeneous distribution of the test item. Amounts of test item, quartz sand and tap water added The treated soil was transferred to the test container and the seeds were sown (see above).
- Controls: A mixture of natural soil and quartz sand
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): quartz sand
- Concentration of vehicle in test medium (stock solution and final test solution): 138 g per treatment
- Evaporation of vehicle before use: Not necessary

Test organisms

1

Species:

Brassica napus

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Rape
- Plant family: Brassicaceae
- Variety: Digger
- Source of seed: KWS Lochow GmbH
- Prior seed treatment/sterilization: No
- Seed storage: The seeds were stored dark at the test facility at room temperature and protected from moisture until use.

2

Species:

Avena sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: Oats
- Plant family: Gramineae
- Variety: Flämingsprofi
- Source of seed: KWS Lochow GmbH
- Prior seed treatment/sterilization: No
- Seed storage: The seeds were stored dark at the test facility at room temperature and protected from moisture until use.

3

Species:

Glycine max (G. soja)

Plant group:

Dicotyledonae (dicots)

Details on test organisms:

- Common name: Soybean
- Plant family: Leguminosae
- Variety: Merlin
- Source of seed: Saatbau Linz
- Prior seed treatment/sterilization: No
- Seed storage: The seeds were stored dark at the test facility at room temperature and protected from moisture until use.

Study design

Test type:

seedling emergence toxicity test

Study type:

laboratory study

Substrate type:

natural soil

Limit test:

yes

Total exposure duration:

21 d

Test conditions

Test temperature:

16 - 25 °C

pH:

5.4 +/- 0.1 (natural soil)

Details on test conditions:

TEST SYSTEM
- Testing facility: Dr.U.Noack-Laboratorien
- Test container (type, material, size): Plastic container (standard flower pots) with a diameter of ca. 12 cm
- Amount of soil: 0.7 kg (dry weight)
- Method of seeding: Test container were filled each with the same amount of treated soil and seeding holes were made with a seedling pistil. One seed was given in each hole and covered with soil.
- No. of seeds per container: 5
- No. of replicates per treatment group: 8
- No. of replicates per control: 8



SOURCE AND PROPERTIES OF SUBSTRATE (if soil): LUFA soil No. 2.2 (batch: Sp2.23708)
- Geographic location: Soil collected by LUFA Speyer (67346 Speyer, Germany)
- Pesticide use history at the collection site: No pesticide use for the last five years.
- Collection procedures: acc. to ISO 10381-6, sampling on 2008-09-18
- Sampling depth (cm): 0 - 20 cm
- Soil texture (if natural soil)
- % sand: 81.4 +/- 1.2
- % silt: 12.2 +/- 0.6
- % clay: 6.4 +/- 0.9
- Soil taxonomic classification: loamy sand
- Soil classification system: German DIN
- Organic carbon (%): 2.16 +/- 0.40
- Maximum water holding capacity (in % dry weigth): 48.2 +/- 5.0
- CEC: 10 +/- 1 mval/100 g
- Pretreatment of soil: soil was stored
- Storage (condition, duration): A 2:1 mixture of natural soil (LUFA 2.2) and quartz sand with a C-content of 1.16 %. The soil has been stored at the test facility at green house conditions until use.


NUTRIENT MEDIUM (if used)
- Description:
Flory 9 hydro (0.5 ‰)
Origin: GRÜNES LANDHAUS,
St. Godehard-Str.23, D-31139 Hildesheim

Nutrient Concentration [%]
Ammonium nitrogen 5
Nitrate nitrogen 10
P2O5 7
Potassium oxide 22
Magnesium oxide 6
Boron 0.03
Iron (Chelate) 0.12
Copper 0.02
Manganese 0.05
Molybdenum 0.005
Zinc 0.01


GROWTH CONDITIONS
- Photoperiod: 16 h
- Light source: Special fluorescent tube Fluora and Daylight (Hoklatherm)
- Light intensity and quality: > 3500 lx
- Day/night temperatures: 22 +/- 10 °C
- Relative humidity (%): 70 +/- 25
- Watering regime and schedules: Bottom watering of the test container with nutrient solution as necessary. The watering interval based on visual observations of soil surface and plants and was checked daily.




EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
During the 21 day observation period the plants were observed every 7 days for number of emerged seedlings, number of dead plants and visual phytotoxic effects. The rating of the treated plants was done in relation to the untreated control plants.
Observations included all variations, either inhibitory or stimulatory, between the treated replicates and the untreated control. Such variations were phytotoxic symptoms (chlorosis, necrosis, wilting), formative effects and growth and development rates.


- Phytotoxicity rating system (if used):

(1) Alteration of form and appearance (deformation)
(1.1) Curling / twisting / sticking or other deformation of the leafs
(1.2) Change of appearance
(1.3) Shoot deformations
(1.4) Wilting
(1.5) Leaf shedding
(1.6) Shedding of the leaf apex
(1.7) Stunted growth
(1.8) Lacking of cotyledons

(2) Discolouration of the leafs
(2.1) Lighter green (2.2) Darker green
(2.3) Chlorosis (2.3.1) Spots
(2.3.2) Whole leaf
(2.3.3) Veins
(2.3.4) Between veins
(2.3.5) Marginal
(2.4) Loss of colour or discolouration

(3) Alteration of the leaf cells
(3.1) Necrosis (3.1.1) Spots
(3.1.2) Marginal
(3.1.3) Apical
(3.1.4) Between veins
(3.1.5) Whole leaf

(E) Dead plant (Number of dead plants)

Assessment of the Effects

- = no effects observed / normal growth
+/- = slight effects
+ = medium effects
++ = strong effects


VEHICLE CONTROL PERFORMED: yes


TEST CONCENTRATIONS

- Range finding study: Not carried out

Nominal and measured concentrations:

1000 mg/kg soil dry weight (nominal)

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Species:

Brassica napus

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Growth and Seed Germination

Effect concentrations 2

Species:

Avena sativa

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Growth and Seed Germination

Effect concentrations 3

Species:

Glycine max (G. soja)

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Growth and Seed Germination

Details on results:

SEEDLING EMERGENCE
- Percent seedling emergence:
Inhibition [%] of Rate of Emergence
Test concentration[mg/kg DW] Rate of Emergence [%]
Oats Inhib. [%] Rape Inhib.[%] Soy-bean Inhib. [%]
Control 88 - 100 - 95 -
1000 90 -3 83 17 95 0




- Plant height:
Inhibition [%] of Shoot Height
Test concentration[mg/kg DW] Shoot Height [cm]
Oats Inhib. [%] Rape Inhib.[%] Soy-bean Inhib. [%]
Control 38.8 - 17.0 - 28.5 -
1000 40.5 -4 17.4 -2 26.9 6




- Fresh weight:
Inhibition [%] of Shoot Fresh Weight
Test concentration[mg/kg DW] Shoot Fresh Weight [mg]
Oats Inhib.[%] Rape Inhib.[%] Soy-bean Inhib. [%]
Control 1194 - 1840 - 3097 -
1000 1383 -16 2334 -27 3059 1






INJURY RATING SYSTEM:
Observation Parameter

(1) Alteration of form and appearance (deformation)
(1.1) Curling / twisting / sticking or other deformation of the leafs
(1.2) Change of appearance
(1.3) Shoot deformations
(1.4) Wilting
(1.5) Leaf shedding
(1.6) Shedding of the leaf apex
(1.7) Stunted growth
(1.8) Lacking of cotyledons

(2) Discolouration of the leafs
(2.1) Lighter green (2.2) Darker green
(2.3) Chlorosis (2.3.1) Spots
(2.3.2) Whole leaf
(2.3.3) Veins
(2.3.4) Between veins
(2.3.5) Marginal
(2.4) Loss of colour
(2.5) Discolouration

(3) Alteration of the leaf cells
(3.1) Necrosis (3.1.1) Spots
(3.1.2) Marginal
(3.1.3) Apical
(3.1.4) Between veins
(3.1.5) Whole leaf

(E) Dead plant (Number of dead plants)


Assessment of the Effects

- = no effects observed / normal growth
+/- = slight effects
+ = medium effects
++ = strong effects

Results with reference substance (positive control):

No reference substance tested

Reported statistics and error estimates:

The percent inhibition of biomass growth (shoot height, shoot fresh weight) and rate of emergence were calculated in relation to the control.

Any other information on results incl. tables

Phytotoxicity rating:

Oats: Visual Observations and Number of Dead Plants on Day 21

Test concentration		Phytotoxic effects
	Phytotoxic	Replicate
[mg/kg DW]	effects*	1	2	3	4	5	6	7	8
Control	(E)	1	-	-	-	-	-	-	-
1000	–	-	-	-	-	-	-	-	-

Rape: Visual Observations and Number of Dead Plants on Day 21

Test concentration		Phytotoxic effects
	Phytotoxic	Replicate
[mg/kg DW]	effects*	1	2	3	4	5	6	7	8
Control	–	-	-	-	-	-	-	-	-
1000	(2.3.1)	-	-	-	-	+/-	-	-	+/-

Soybean: Visual Observations and Number of Dead Plants on Day 21

Test concentration		Phytotoxic effects
	Phytotoxic	Replicate
[mg/kg DW]	effects*	1	2	3	4	5	6	7	8
Control	–	-	-	-	-	-	-	-	-
1000	(1.8)	-	-	-	-	-	1	-	-

Inhibition [%] of Shoot Height

Test concentration [mg/kg DW]		Shoot Height [cm]
		Oats		Inhib. [%]		Rape		Inhib. [%]		Soy-bean		Inhib. [%]
Control		38.8		-		17.0		-		28.5		-
1000		40.5		-4		17.4		-2		26.9		6

Inhibition [%] of Shoot Fresh Weight

Test concentration [mg/kg DW]	Shoot Fresh Weight [mg]
	Oats	Inhib. [%]	Rape	Inhib. [%]	Soy-bean	Inhib. [%]
Control	1194	-	1840	-	3097	-
1000	1383	-16	2334	-27	3059	1

Inhib. = Inhibition                      negative values = promoted growth

Inhibition [%] of Rate of Emergence

Test concentration [mg/kg DW]	Rate of Emergence [%]
	Oats	Inhib. [%]	Rape	Inhib. [%]	Soy-bean	Inhib. [%]
Control	88	-	100	-	95	-
1000	90	-3	83	17	95	0

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The test item Pigment Red 112 is not toxic at concentration After 21 days no test item related phytotoxic effects were observed for the tested plant species

Executive summary:

The phytotoxicity of the test item Pigment Red 112 to three terrestrial plant species was determined in a seedling emergence test over a period of 21 days according to OECD 208. The test was conducted with the nominal test item concentration 1000 mg/kg soil dry weight (DW). Test systems were one monocotyledon, oats (Gramineae), and two dicotyledons, rape (Brassicaceae) and soybean (Leguminosae). The toxic effects of the test item were determined on day 7, 14 and 21 by visual observations (number of emerged seedlings and phytotoxic effects) and on day 21 by shoot height and shoot fresh weight determination. The inhibition of shoot height, shoot fresh weight and rate of emergence was calculated in relation to the untreated control. The test item did not cause any effects > 25 % on the shoot height, the shoot fresh weight and the rate of emergence of the tested plant species at the tested concentration. After 21 days no test item related phytotoxic effects were observed for the tested plant species.

Table 1:    Inhibition of Shoot Height, Shoot Fresh Weight and Rate of Emergence
in Comparison to the Control

	Inhibition [%]
Species	Shoot height	Shoot fresh weight	Rate of Emergence
Oats	-4	-16	-3
Rape	-2	-27	17
Soybean	6	1	0

The test item Pigment Red 112 is not toxic at concentration ≤ 1000 mg/kg soil dry weight to the tested plant species.