Quaternary ammonium compounds, C12-14-alkyl(hydroxyethyl)dimethyl, chlorides

Toxicity to terrestrial plants

Administrative data

Endpoint:

toxicity to terrestrial plants: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993-07-16 to 1993-08-21

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This study was conducted using methods according to OECD guideline 208. Study was well documented and GLP.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

N/A

GLP compliance:

yes

Remarks:

stated, no certificate included

Test material

Test material information

Constituent 1

Reference substance name:

[see concordance under 1.1. of the CSR]

IUPAC Name:

[see concordance under 1.1. of the CSR]

Details on test material:

- Name of test material (as cited in study report): C12/14 KDB (E-4408.01); C12/14 Alkyl dimethyl hydroxyethyl ammonium chloride
- Molecular formula (if other than submission substance): N/A
- Molecular weight (if other than submission substance): N/A
- Smiles notation (if other than submission substance): N/A
- InChl (if other than submission substance): N/A
- Structural formula attached as image file (if other than submission substance): see Fig. N/A
- Substance type: Active
- Physical state: white, transparent, liquid

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
PHYSICO-CHEMICAL PROPERTIES
- Vapour pressure: .NK
- Water solubility (under test conditions): good
- Henry's law constant: N/A
- log Pow: N/A
- pKa: N/A
- UV absorption: N/A
- pH dependance on stability: 7+-1.5
- Soil sorption behaviour: N/A
- pKa: N/A
- Stability in water: yes
- Stability in light: yes


OTHER PROPERTIES (if relevant for this endpoint)
- Biodegradability: N/A
- Other: N/A

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations: samples from the control and all stock solutions (37830.9, 12105.8, 3783.7, 1210.9, 378.4 and 121.0 mg a.i. per litre) were taken at t=0 (newly prepared solutions).
- Sampling method: Samples of ca. 100 ml were taken in glass bottles. Aliquots of 3 ml formaldehyde (37% v/v) and 0.1 ml of a nonionic stock were added to each glass bottle before 100 ml of each sample was added. The analysis of these test solutions was arranged by Procter and Gamble ETC.
- Sample storage conditions before analysis: The bottles were stored in a refrigerator until transfer to Procter and Gamble.

Test substrate

Vehicle:

no

Details on preparation and application of test substrate:

- Method of application to filter paper (if used): N/A
- Method of mixing into soil (if used): Quantities of 23644.3, 7566.1, 2364.8, 756.8, 236.5, and 75.6 mg of the test substance were accurately weighed out and dissolved in 250 ml of demineralized water respectively, resulting in stock solutions of 37830.9, 12105.8, 3783.7, 1210.9, 378.4 and 121.0 mg a.i. per litre respectively. An aliquot of 50 ml of a stock solution of 62.8 g K2HPO4 per liter (added to obtain an optimum growth) and 50 ml of each stock solution, prepared as mentioned above, were added respectively to 2000 g of damp soil (1891.5 g of dry soil and 108.5 g water) and thoroughly mixed with a household mixer to achieve test concentrations of 1000, 320, 100, 32, 10, and 3.2 mg a.i. per kg of dry soil.
- Method of application to soil surface (if used): N/A
- Controls: The control was prepared in a similar manner by adding 50 ml of the stock solution of K2HPO4 and 50 ml of demineralized water to 2000 g of damp soil.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): N/A
- Concentration of vehicle in test medium (stock solution and final test solution): N/A
- Evaporation of vehicle before use: N/A

Test organisms

Species:

Lactuca sativa

Plant group:

Monocotyledonae (monocots)

Details on test organisms:

- Common name: lettuce
- Plant family: Asteraceae
- Variety: Ravel RZ
- Source of seed: Rijk Zwaan (De Lier, the Netherlands)
- Prior seed treatment/sterilization: N/A
- Historical germination of seed (germination of seed lot tested): N/A
- Seed storage: N/A

Study design

Test type:

other: seedling emergence and growth

Study type:

laboratory study

Substrate type:

other: semi-natural

Limit test:

no

Total exposure duration:

18 d

Remarks:

N/A

Post exposure observation period:

N/A

Test conditions

Test temperature:

21-26 degree C

pH:

pH at start: 7.2
pH at end: 7.7

Moisture:

25% (based on dry constituents)

Details on test conditions:

TEST SYSTEM
- Testing facility: TNO
- Test container (type, material, size): The test vessels were circular plastic cups (food quality) with a maximum diameter of about 9.3 cm and a height of about 6.0 cm. The cups were about 2/3 filled with soil.
- Amount of soil: 355 g
- Method of seeding: The seeds of Lactuca sativa (lettuce) were sown on top of the soil. No water was added after the sowing of Lactuca sativa (lettuce).
- No. of seeds per container: ~ 10 seeds were sown in each of the cups
- No. of plants (retained after thinning): 5 seedlings
- No. of replicates per treatment group: 4
- No. of replicates per control: 4
- No. of replicates per vehicle control: N/A


SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Heerewaarden (the Netherlands) agricultural soil from an orchard mixed with coarse sand (1:1).
- Pesticide use history at the collection site: N/A
- Collection procedures: N/A
- Sampling depth (cm): N/A
- Soil texture (if natural soil)
- % sand: 83.7%
- % silt: 10.1 %
- % clay: N/A
- Soil taxonomic classification: N/A
- Soil classification system: N/A
- Composition (if artificial substrate): N/A
- Organic carbon (%): 1:1%
- Maximum water holding capacity (in % dry weight): N/A
- CEC: N/A
- Pretreatment of soil: 1.66 g K2HPO4 added per kg dry soil.
- Storage (condition, duration): N/A


NUTRIENT MEDIUM (if used)
- Description: Semi-natural soil: A quantity of 1.66 g K2HPO4 was added per kg of dry soil to assure good growth conditions.
A quantity of 62.8 g K2HPO4 was added to an aliquot of 50 ml of a stock solution to obtain optimum growth.



GROWTH CONDITIONS
- Photoperiod: 16 h light - 8 h dark
- Light source: N/A
- Light intensity and quality: 6800 lux (lettuce)
- Day/night temperatures: 21.5 degree C (dark period) and 25.5 degree C (light period).
- Relative humidity (%): N/A
- Wind velocity: N/A
- Air exchange rate: N/A
- Watering regime and schedules: N/A
- Water source/type: demineralized water
- Volume applied: N/A
- Interval of applications: daily addition of demineralized water to bring moisture content back to the initial level.
- Method of application: At the start of the tests, the soil had a moisture content of about 25% (based on dry constituents). About 10 seeds were sown in each of the cups and covered with a glass plate. The space between the glass plate and top of the soil was about 2 cm. The plate was removed after the seeds had emerged and before they grew tall enough to touch it. All emergence of seedlings was recorded after 3 days. Five healthy-looking seedlings from each treatment were left to grow for another 14 days, and weighed at the end of the test. After the glass plates had been removed, the cups were weighed daily, and demineralized water was added to the soil to bring the moisture content back to the initial level ( the weight of the plants in the cup was ignored and to compensate for the weight of the plastic cup, the balance was tared using another plastic cup).
- Any pest control method/fertilization (if used): N/A



ACCLIMATION PERIOD: N/A


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : The endpoints were emergence of the seedlings, growth (wet-weight at the end of the test), survival of seedlings and visual appearance of the plants.


- Phytotoxicity rating system (if used): Appearance, height, stem thickness.


VEHICLE CONTROL PERFORMED: N/A


TEST CONCENTRATIONS
- Spacing factor for test concentrations: N/A
- Justification for using less concentration than requested by guideline: N/A
- Range finding study: No
- Test concentrations: N/A
- Results used to determine the conditions for the definitive study: N/A

Nominal and measured concentrations:

Nominal concentrations: 0, 3.2, 10, 32, 100, 320, and 1000 mg of a.i. per kg of dry soil.
Measured concentrations: Samples of the stock solutions were taken, preserved, and sent to the sponsor for analysis, but the results are not available in this study report.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

EC10

Effect conc.:

10.3 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Remarks:

wet-weight

Remarks on result:

other: 6.0 - 17.5

Effect concentrations 2

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

other: EC20

Effect conc.:

15.6 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Remarks:

wet-weight

Remarks on result:

other: 10.0 - 24.2

Effect concentrations 3

Species:

Lactuca sativa

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: emergence

Effect concentrations 4

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: survival of seedlings

Effect concentrations 5

Species:

Lactuca sativa

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

3.2 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

growth

Remarks:

wet-weight

Details on results:

SEED GERMINATION
- Percent seed germination: N/A
- Root length: N/A
- Root discolouration/malformation: N/A
- Other effects: N/A
- Results with reference substance valid? N/A


SEEDLING EMERGENCE
- Percent seedling emergence: see attached table 1
- Percent survival: 100%
- Plant height: N/A
- Dry weight: N/A
- Abnormal seed development or appearance: see attached table 1
- Lesions: N/A
- Swelling: N/A
- Loss of turgour: N/A
- Discoloration: no leaf damage (e.g. yellow or brown spots) to any of the plants in any of the concentrations tested was seen during the exposure period.
- Unusual leaf/plant shape or size: see attached table 1
- Other effects: N/A


VEGETATIVE VIGOUR
- Percent seed germination: N/A
- Plant height: N/A
- Dry weight: N/A
- Abnormal seed development or appearance: N/A
- Lesions: N/A
- Swelling: N/A
- Loss of turgour: N/A
- Discoloration: N/A
- Unusual leaf/plant shape or size: N/A
- Dead plants: 0
- Other effects: N/A


INJURY RATING SYSTEM: Adverse effects on plant appearance, height and growth were observed by day 7 at doses >= 10 mg/kg a.i.

Results with reference substance (positive control):

N/A

Reported statistics and error estimates:

NOECs were calculated using JMP 7.0 Statistical Software. EC10s and EC20s were calculated using SAS v. 9.1. The results from these calculations are presented in this robust summary.

In the original study report, the EC50 (growth) values were calculated using an iterative maximum likelihood estimation procedure and assuming a relationship between wet-weight (W) at the end of the exposure period and exposure concentration (C) of the form.

To determine the NOEC for emergence and mortality, the dates on emergence of seedlings and the mortality dates of each concentration were compared pair-wise with those in the control using a binomial test. A significance level of 5% was used. Effects on emergence are possible in the form of postponement and of complete prevention.

To determine the NOEC for wet-weight at the end of the test, a multiple comparison was made between the wet-weight of each plant at each concentration and the wet-weight of the plants in the control using a two tailed Dunnett-test. It was assumed that the plants in each cup were growing independently of each other. A significance level of 5% was used.

Any other information on results incl. tables

Table 1. Effect of KDB on Lettuce (Lactuca sativa)

Nominal Conc. (mg a.i./kg dry wt soil)	Emergence (# out of 40 total)	Mortality of Seedlings (# out of 20 total)	Growth (average wet weight)
0	39	0	2.44
3.2	37	0	2.50
10	39	0	2.15
32	36	1	1.51 *
100	37	0	0.19*
320	36	0	0.04 *
1000	37	1	0.01 *

* Significantly different from control (p=0.95). 20 plants/test concentration.

Table 2. Summary of Effects of KDB on Oats and Lettuce (study # 41971)

Parameter	Effect	mg a.i./kg dry wt soil (nominal conc.)
		Oats (Avena sativa)	Lettuce (Lactuca sativa)
Emergence	NOEC	100	>1000
	EC10	155	---
	EC20	1090	---
Survival of Seedlings	NOEC	>1000	>1000
Growth (wet wt.)	NOEC	3.2	3.2
	EC10	6.1	10.3
	EC20	15.7	15.6

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The effect of KDB to the plant species Lactuca sativa (lettuce) was determined according to OECD Guideline No. 208. The EC10 for growth (wet weight) was 10.3 mg ai/kg soil dry wt.

Executive summary:

The effect of C12 -14 dimethyl hydroxyethyl ammonium chloride (KDB) on the plant species Lactuca sativa (lettuce) was determined according OECD Guideline No. 208. The parameters measured included emergence of the seedlings, growth (wet-weight at the end of the test), and survival of seedlings. Nominal test concentrations were 0, 3.2, 10, 32, 100, 320, and 1000 mg a.i./kg dry soil.

In each concentration of the test substance, 4x5 plants were grown in a mixture of agricultural soil and coarse sand, supplemented with potassium and phosphorus. Lactuca sativa (lettuce) was exposed for 18 days. Emergence of more than 50% of Lactuca sativa (lettuce) seedlings in the control occurred within 4 days. After emergence the plants were further exposed for a growth period of 14 days.

Emergence of Lactuca sativa (lettuce) was slightly slower than for Avena sativa (oats), the other species tested in this study. See summary referenced below. Most seeds were emerged at day 4. For Lactuca sativa (lettuce) no significant reduction of emergence was observed at the highest test concentration (1000 mg a.i..kg-1), 37 out of the 40 seeds emerged.

There was no significant effect on seedling survival at any test concentration.

The results of the study were as follows:

EC10 (growth) = 10.3 mg ai/kg dry soil,

NOEC (emergence) = >1000 mg ai/kg dry soil.

This toxicity study satisfies the guideline requirements for the OECD 208 Terrestrial Plants Test: Seedling Emergence and Growth.